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Sample records for bifidobacterium lactis hn019

  1. Adhesion and immunomodulatory effects of Bifidobacterium lactis HN019 on intestinal epithelial cells INT-407

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    AIM:To elucidate the adherence and immunomodulatory properties of a probiotic strain Bifidobacterium lactis(B.lactis) HN019.METHODS:Adhesion assays of B.lactis HN019 and Salmonella typhimurium(S.typhimurium) ATCC 14028 to INT-407 cells were carried out by detecting copies of species-specific genes with real-time polymerase chain reaction.Morphological study was further conducted by transmission electron microscopy.Interleukin-1β(IL-1β),interleukin-8,and tumor necrosis factor-α(TNF-α) gene expression were as...

  2. Dose-response effect of Bifidobacterium lactis HN019 on whole gut transit time and functional gastrointestinal symptoms in adults

    OpenAIRE

    2011-01-01

    Objective. To assess the impact of Bifidobacterium lactis HN019 supplementation on whole gut transit time (WGTT) and frequency of functional gastrointestinal (GI) symptoms in adults. Material and methods. We randomized 100 subjects (mean age: 44 years; 64% female) with functional GI symptoms to consume a proprietary probiotic strain, B. lactis HN019 (Fonterra Research Centre, Palmerston North, New Zealand), at daily doses of 17.2 billion colony forming units (CFU) (high dose; n = 33), 1.8 bil...

  3. Promotility Action of the Probiotic Bifidobacterium lactis HN019 Extract Compared with Prucalopride in Isolated Rat Large Intestine

    Science.gov (United States)

    Dalziel, Julie E.; Anderson, Rachel C.; Peters, Jason S.; Lynch, Amy T.; Spencer, Nick J.; Dekker, James; Roy, Nicole C.

    2017-01-01

    Attention is increasingly being focussed on probiotics as potential agents to restore or improve gastrointestinal (GI) transit. Determining mechanism of action would support robust health claims. The probiotic bacterium Bifidobacterium lactis HN019 reduces transit time, but its mechanisms of action and effects on motility patterns are poorly understood. The aim of this study was to investigate changes in GI motility induced by an extract of HN019 on distinct patterns of colonic motility in isolated rat large intestine, compared with a known promotility modulator, prucalopride. The large intestines from male Sprague Dawley rats (3–6 months) were perfused with Kreb's buffer at 37°C in an oxygenated tissue bath. Isometric force transducers recorded changes in circular muscle activity at four independent locations assessing contractile propagation between the proximal colon and the rectum. HN019 extract was perfused through the tissue bath and differences in tension and frequency quantified relative to pre-treatment controls. Prucalopride (1 μM) increased the frequency of propagating contractions (by 75 ± 26%) in the majority of preparations studied (10/12), concurrently decreasing the frequency of non-propagating contractions (by 50 ± 11%). HN019 extract had no effect on contractile activity during exposure (n = 8). However, following wash out, contraction amplitude of propagating contractions increased (by 55 ± 18%) in the distal colon, while the frequency of non-propagating proximal contractions decreased by 57 ± 7%. The prokinetic action of prucalopride increased the frequency of synchronous contractions along the length of colon, likely explaining increased colonic rate of transit in vivo. HN019 extract modified motility patterns in a different manner by promoting propagating contractile amplitude and inhibiting non-propagations, also demonstrating prokinetic activity consistent with the reduction of constipation by B. lactis HN019 in humans. PMID:28184185

  4. Survival of Lactobacillus acidophilus NCFM® and Bifidobacterium lactis HN019 encapsulated in chocolate during in vitro simulated passage of the upper gastrointestinal tract

    DEFF Research Database (Denmark)

    Klindt-Toldam, Stine; Larsen, Susanne K.; Saaby, Lasse

    2016-01-01

    Probiotic microorganisms are exposed to different types of stress both before and after consumption which strongly influences probiotic survival. Here the viability of 2 different probiotic bacteria incorporated in different types of chocolate was investigated during storage and passage of the up......Probiotic microorganisms are exposed to different types of stress both before and after consumption which strongly influences probiotic survival. Here the viability of 2 different probiotic bacteria incorporated in different types of chocolate was investigated during storage and passage...... of the upper gastro-intestinal tract using both a static and a dynamic gastric in vitro model. Lactobacillus acidophilus NCFM® and Bifidobacterium lactis HN019 equalling a total concentration of 2 × 108, 2 × 109 and 2 × 1010 CFU/g chocolate were added to samples of milk chocolate, 57% and 72% dark chocolate...... dairy and juice based probiotic products, chocolate was an excellent carrier for probiotic delivery, because of very good survival of probiotics during simulated passage of the upper GI tract. The viability of B. lactis was slightly higher than Lb. acidophilus and survival rates were >6.5 log CFU...

  5. Transcriptional analysis of oligosaccharide utilization by Bifidobacterium lactis Bl-04

    DEFF Research Database (Denmark)

    Andersen, Joakim Mark; Barrangou, Rodolphe; Abou Hachem, Maher

    2013-01-01

    transcriptomes of Bifidobacterium animalis subsp. lactis Bl-04, induced by 11 potential prebiotic oligosaccharides were analyzed to identify the genetic loci involved in the uptake and catabolism of α- and β-linked hexoses, and β-xylosides. RESULTS: The overall transcriptome was modulated dependent on the type...... in the breadth and selectivity of prebiotic utilization by bifidobacteria. CONCLUSION: This study identified the differential gene expression for utilization of potential prebiotics highlighting the extensive capabilities of Bifidobacterium lactis Bl-04 to utilize oligosaccharides. Results provide insights...

  6. Insights into physiological traits of Bifidobacterium animalis subsp. lactis BB-12 through membrane proteome analysis

    DEFF Research Database (Denmark)

    Gilad, Ofir; Hjernø, Karin; Østerlund, Eva Christina

    2012-01-01

    Bifidobacterium animalis subsp. lactis BB-12 is a widely used probiotic strain associated with a variety of health-promoting traits. There is, however, only limited knowledge available regarding the membrane proteome and the proteins involved in oligosaccharide transport in BB-12. We applied two...

  7. Bifidobacterium lactis attenuates onset of inflammation in a murine model of colitis

    Institute of Scientific and Technical Information of China (English)

    David Philippe; Stéphanie Blum; Laurent Favre; Francis Foata; Oskar Adolfsson; Genevieve Perruisseau-Carrier; Karine Vidal; Gloria Reuteler; Johanna Dayer-Schneider; Christoph Mueller

    2011-01-01

    AIM: To assess the anti-inflammatory effect of the probiotic Bifidobacterium lactis (B. lactis ) in an adoptive transfer model of colitis. METHODS: Donor and recipient mice received either B. lactis or bacterial culture medium as control (deMan Rogosa Sharpe) in drinking water for one week prior to transfer of a mix of naive and regulatory T cells until sacrifice. RESULTS: All recipient mice developed signs of colonic inflammation, but a significant reduction of weight loss was observed in B. lactis -fed recipient mice compared to control mice. Moreover, a trend toward a diminution of mucosal thickness and attenuated epithelial damage was revealed. Colonic expression of pro-inflammatory and T cell markers was significantly reduced in B. lactis - fed recipient mice compared to controls. Concomitantly, forkhead box protein 3, a marker of regulatory T cells, was significantly up-regulated by B. lactis . CONCLUSION: Daily oral administration of B. lactis was able to reduce inflammatory and T cells mediators and to promote regulatory T cells specific markers in a mouse model of colitis.

  8. Impact of Bifidobacterium lactis supplementation on fecal microbiota in infants delivered vaginally compared to Caesarean section

    Directory of Open Access Journals (Sweden)

    Tetty Yuniaty

    2013-03-01

    Full Text Available AbstractBackground It has been reported that infants born by Caesarean section have altered gut microbiota, with lower numbers of bifidobacteria and Bacteroides, compared to that of infants who were delivered vaginally. Probiotic supplementation has been reported to have beneficial effects on the immune response, generally in relation to allergies.Objective To assess the effect of Bifidobacterium lactis (B. lactis supplementation on the presence of B. lactis and bifidobacteria counts in stool of infants during the first 2 months of life.Methods We conducted an observational study of 122 healthy, breast-fed infants delivered vaginally or by Caesarean section. Infants assigned to the test group received breast milk and formula supplemented with the B. lactis probiotics. Infants in the control group received breast milk and formula without probiotics. The presence of B. lactis and stool bifidobacteria counts were determined at 1 month and 2 months of age. Growth, morbidity, serum immune markers, and stool immunoglobulin (Ig A were also assessed.Results B. lactis was more frequently detected in the stool of infants who received breast milk and probiotic-supplemented formula than in stool of infants who received breast milk and non-supplemented formula, both at 1 month and 2 months of age (OR 1,263; 95%CI 11 to 151,030; P=0.003. Of infants who received probiotic-supplemented formula, B. lactis was detected in 80% of those delivered by Caesarean section and in 38% of those delivered vaginally, at the 1-month mark. In infants delivered by Caesarean section, the mean stool bifidobacteria level at 1 month was significantly higher in the probiotic-supplemented group compared to that of the non-supplemented group (P=0.021.Conclusion Eearly bifidobacteria supplementation of infants, particularly those delivered by Caesarean section, is associated with higher levels of stool bifidobacteria. Anthropometric data suggests beneficial effects of bifidobacteria

  9. Impact of Bifidobacterium lactis supplementation on fecal microbiota in infants delivered vaginally compared to Caesarean section

    Directory of Open Access Journals (Sweden)

    Tetty Yuniaty

    2013-03-01

    Full Text Available Background It has been reported that infants born by Caesarean section have altered gut microbiota, with lower numbers of bifidobacteria and Bacteroides, compared to that of infants who were delivered vaginally. Probiotic supplementation has been reported to have beneficial effects on the immune response, generally in relation to allergies. Objective To assess the effect of Bifidobacterium lactis (B. lactis supplementation on the presence of B. lactis and bifidobacteria counts in stool of infants during the first 2 months of life. Methods We conducted an observational study of 122 healthy, breast-fed infants delivered vaginally or by Caesarean section. Infants assigned to the test group received breast milk and formula supplemented with the B. lactis probiotics. Infants in the control group received breast milk and formula without probiotics. The presence of B. lactis and stool bifidobacteria counts were determined at 1 month and 2 months of age. Growth, morbidity, serum immune markers, and stool immunoglobulin (Ig A were also assessed. Results B. lactis was more frequently detected in the stool of infants who received breast milk and probiotic-supplemented formula than in stool of infants who received breast milk and non-supplemented formula, both at 1 month and 2 months of age (OR 1,263; 95%CI 11 to 151,030; P=0.003. Of infants who received probiotic-supplemented formula, B. lactis was detected in 80% of those delivered by Caesarean section and in 38% of those delivered vaginally, at the 1-month mark. In infants delivered by Caesarean section, the mean stool bifidobacteria level at 1 month was significantly higher in the probiotic-supplemented group compared to that of the non-supplemented group (P=0.021. Conclusion Eearly bifidobacteria supplementation of infants, particularly those delivered by Caesarean section, is associated with higher levels of stool bifidobacteria. Anthropometric data suggests beneficial effects of bifidobacteria

  10. Prebiotic Effects of Agave salmiana Fructans in Lactobacillus acidophilus and Bifidobacterium lactis Cultures.

    Science.gov (United States)

    Castro-Zavala, Adriana; Juárez-Flores, Bertha I; Pinos-Rodríguez, Juan M; Delgado-Portales, Rosa E; Aguirre-Rivera, Juan R; Alcocer-Gouyonnet, Francisco

    2015-11-01

    Agave salmiana is a fructan rich species that is widely distributed in Mexico. The aim of this investigation was to extract the fructans of A. salmiana and evaluate their prebiotic effect in 48 hours in vitro cultures of Bifidobacterium lactis and Lactobacillus acidophilus and to compare this effect with other available fructan sources. A significant difference in pH, optical density and biomass was found in the cultures depending on the source of fructans and the type of bacteria. It was possible to determine a dose-response effect of the A. salmiana fructans and the growth of the studied strains.

  11. EFFECT OF THE CONSUMPTION OF A CHEESE ENRICHED WITH PROBIOTIC ORGANISMS (BIFIDOBACTERIUM LACTIS BI-07 IN IMPROVING SYMPTOMS OF CONSTIPATION

    Directory of Open Access Journals (Sweden)

    Diane Cassia FAVRETTO

    2013-09-01

    Full Text Available Context Constipation is a very common symptom in the general population. One way of non-pharmacological treatment of constipation is through the addition of probiotics to food. Obectives The aim of this study was to evaluate de effect of the consumption of a fresh cheese, enriched with Bifidobacterium lactis Bi-07 on the symptoms of constipated women. Methods A randomized controlled trial, carried out in the Basic Health Units of Guaporé's City – RS/Brazil, between january and may 2012, with 30 constipated women. The patients were randomized into two groups whom received, for 30 days, 30 g of fresh cheese enriched with Bifidobacterium lactis Bi-07 (n = 15 or regular fresh cheese (n = 15. Constipation symptoms were evaluated according to ROMA III Consensus, before and after the nutritional intervention. Also, data of clinical and anthropometric characteristics of the individuals were collected. Accepted level of significance 5% (P≤0,05. Results The medium age of the studied population was 37,5±14,4 years in the intervention group and 40,8±12,8 years in the control group. After 30 days we observed that the ingestion of fresh cheese enriched with Bifidobacterium lactis Bi-07 promoted benefic effects on the symptoms of strength to evacuate. Conclusion The consumption of 30g/day of a fresh cheese enriched with Bifidobacterium lactis Bi-07 has beneficial effects on constipation symptoms.

  12. Stimulation of the growth of two probiotic bacteria, Lactobacillus Acidophilus NCFM and Bifidobacterium Lactis BL-04, by selected prebiotic canditates

    DEFF Research Database (Denmark)

    van Zanten, Gabriella Christina; Knudsen, Anne; Bandsholm, O.

    2009-01-01

    Prebiotics have been shown to support the growth of probiotic bacteria thereby having a beneficial effect on human health. The aim of this work was to evaluate selected and structurally different carbohydrate prebiotic candidate by measuring their capability to stimulate the growth of the two pro...... probiotic bacteria Lactobacillus acdophilus NCFM and Bifidobacterium lactis BL-04....

  13. Analysis of the human intestinal epithelial cell transcriptional response to Lactobacillus acidophilus, Lactobacillus salivarius, Bifidobacterium lactis and Escherichia coli

    DEFF Research Database (Denmark)

    Putaala, H; Barrangou, R; Leyer, G J

    2010-01-01

    a comparative analysis of the global in vitro transcriptional response of human intestinal epithelial cells to Lactobacillus acidophilus NCFM™, Lactobacillus salivarius Ls-33, Bifidobacterium animalis subsp. lactis 420, and enterohaemorrhagic Escherichia coli O157:H7 (EHEC). Interestingly, L. salivarius Ls-33...

  14. Stimulation of the growth of two probiotic bacteria, Lactobacillus Acidophilus NCFM and Bifidobacterium Lactis BL-04, by selected prebiotic canditates

    DEFF Research Database (Denmark)

    van Zanten, Gabriella Christina; Knudsen, Anne; Bandsholm, O.;

    2009-01-01

    Prebiotics have been shown to support the growth of probiotic bacteria thereby having a beneficial effect on human health. The aim of this work was to evaluate selected and structurally different carbohydrate prebiotic candidate by measuring their capability to stimulate the growth of the two...... probiotic bacteria Lactobacillus acdophilus NCFM and Bifidobacterium lactis BL-04....

  15. Production of bacteriocin-like inhibitory substance by Bifidobacterium lactis in skim milk supplemented with additives.

    Science.gov (United States)

    Martinez, Fabio Andres Castillo; Domínguez, José Manuel; Converti, Attilio; Oliveira, Ricardo Pinheiro de Souza

    2015-08-01

    Bacteriocins are natural compounds used as food biopreservatives instead of chemical preservatives. Bifidobacterium animalis subsp. lactis (Bifid. lactis) was shown to produce a bacteriocin-like inhibitory substance (BLIS) able to inhibit the growth of Listeria monocytogenes selected as an indicator microorganism. To enhance this production by the strain Bifid. lactis BL 04, skim milk (SM) was used as a fermentation medium either in the presence or in the absence of yeast extract, Tween 80 or inulin as stimulating additives, and the results in terms of bacterial growth and BLIS production were compared with those obtained in a traditional high cost complex medium such as Man, Rogosa and Sharpe (MRS). To this purpose, all the cultivations were carried out in flasks at 200 rpm under anaerobic conditions ensured by a nitrogen flowrate of 1.0 L/min for 48 h, and BLIS production was quantified by means of a modified agar diffusion assay at low values of both temperature and concentration of List. monocytogenes. Although all these ingredients were shown to exert positive influence on BLIS production in both media, yeast extract and SM were by far the best ingredient and the best medium, respectively, allowing for a BLIS production at the late exponential phase of 2000 AU/ml.

  16. Effect of composite yogurt enriched with acacia fiber and Bifidobacterium lactis

    Institute of Scientific and Technical Information of China (English)

    Yang Won Min; Sang Un Park; Yeon Sil Jang; Young-Ho Kim; Poong-Lyul Rhee; Seo Hyun Ko; Nami Joo

    2012-01-01

    AIM:To investigate whether composite yogurt with acacia dietary fiber and Bifidobacterium lactis (B.lactis)has additive effects in irritable bowel syndrome (IBS).METHODS:A total of 130 patients were randomly allocated to consume,twice daily for 8 wk,either the composite yogurt or the control product.The composite yogurt contained acacia dietary fiber and high-dose B.lactis together with two classic yogurt starter cultures.Patients were evaluated using the visual analog scale via a structured questionnaire administered at baseline and after treatment.RESULTS:Improvements in bowel habit satisfaction and overall IBS symptoms from baseline were significantly higher in the test group than in the control group (27.16 vs 15.51,P =0.010,64.2 ± 17.0 vs 50.4 ± 20.5,P < 0.001; respectively).In constipation-predominant IBS,improvement in overall IBS symptoms was significantly higher in the test group than in the control group (72.4 ± 18.4 vs 50.0 ± 21.8,P < 0.001).In patients with diarrhea-predominant IBS,improvement in bowel habit satisfaction from baseline was significantly higher in the test group than in the control group (32.90 vs 7.81,P =0.006).CONCLUSION:Our data suggest that composite yogurt enriched with acacia fiber and B.lactis has greater therapeutic effects in patients with IBS than standard yogurt.

  17. A randomised double-blind placebo-controlled trial with Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BB-12 for maintenance of remission in ulcerative colitis

    DEFF Research Database (Denmark)

    Nordgaard, Inge; Rumessen, Jüri Johs.; Wildt, Signe

    2011-01-01

    To investigate the clinical effect of treatment with Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BB-12 (Probio-Tec AB-25) to maintain remission in patients with ulcerative colitis.......To investigate the clinical effect of treatment with Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BB-12 (Probio-Tec AB-25) to maintain remission in patients with ulcerative colitis....

  18. Microencapsulation of Bifidobacterium animalis subsp. lactis and Lactobacillus acidophilus in cocoa butter using spray chilling technology.

    Science.gov (United States)

    Pedroso, D L; Dogenski, M; Thomazini, M; Heinemann, R J B; Favaro-Trindade, C S

    2013-01-01

    In the present study, the cells of Bifidobacterium animalis subsp. lactis (BI-01) and Lactobacillus acidophilus (LAC-04) were encapsulated in cocoa butter using spray-chilling technology. Survival assays were conducted to evaluate the resistance of the probiotics to the spray-chilling process, their resistance to the simulated gastric and intestinal fluids (SGF and SIF), and their stability during 90 days of storage. The viability of the cells was not affected by microencapsulation. The free and encapsulated cells of B. animalis subsp. lactis were resistant to both SGF and SIF. The micro-encapsulated cells of L. acidophilus were more resistant to SGF and SIF than the free cells; the viability of the encapsulated cells was enhanced by 67%, while the free cells reached the detection limit of the method (10(3) CFU/g). The encapsulated probiotics were unstable when they were stored at 20 °C. The population of encapsulated L. acidophilus decreased drastically when they were stored at 7 °C; only 20% of cells were viable after 90 days of storage. The percentage of viable cells of the encapsulated B. animalis subsp.lactis, however, was 72% after the same period of storage. Promising results were obtained when the microparticles were stored at -18 °C; the freeze granted 90 days of shelf life to the encapsulated cells. These results suggest that the spray-chilling process using cocoa butter as carrier protects L. acidophilus from gastrointestinal fluids. However, the viability of the cells during storage must be improved.

  19. Microencapsulation of Bifidobacterium animalis subsp. lactis and Lactobacillus acidophilus in cocoa butter using spray chilling technology

    Directory of Open Access Journals (Sweden)

    D.L. Pedroso

    2013-09-01

    Full Text Available In the present study, the cells of Bifidobacterium animalis subsp. lactis (BI-01 and Lactobacillus acidophilus (LAC-04 were encapsulated in cocoa butter using spray-chilling technology. Survival assays were conducted to evaluate the resistance of the probiotics to the spray-chilling process, their resistance to the simulated gastric and intestinal fluids (SGF and SIF, and their stability during 90 days of storage. The viability of the cells was not affected by microencapsulation. The free and encapsulated cells of B. animalis subsp. lactis were resistant to both SGF and SIF. The micro-encapsulated cells of L. acidophilus were more resistant to SGF and SIF than the free cells; the viability of the encapsulated cells was enhanced by 67%, while the free cells reached the detection limit of the method (10³ CFU/g. The encapsulated probiotics were unstable when they were stored at 20 °C. The population of encapsulated L. acidophilus decreased drastically when they were stored at 7 °C; only 20% of cells were viable after 90 days of storage. The percentage of viable cells of the encapsulated B. animalis subsp.lactis, however, was 72% after the same period of storage. Promising results were obtained when the microparticles were stored at -18 °C; the freeze granted 90 days of shelf life to the encapsulated cells. These results suggest that the spray-chilling process using cocoa butter as carrier protects L. acidophilus from gastrointestinal fluids. However, the viability of the cells during storage must be improved.

  20. The influence of different polymers on viability of Bifidobacterium lactis 300b during encapsulation, freeze-drying and storage

    OpenAIRE

    Pop, Oana Lelia; Brandau, Thorsten; Schwinn, Jens; Dan Cristian VODNAR; Carmen SOCACIU

    2014-01-01

    Seven different types of natural polymers namely hydroxypropyl methylcellulose (HPMC), sodium-carboxymethyl cellulose (Na-CMC), microcrystalline cellulose (MCC), starch BR-07, starch BR-08, dextrin and pullulan were used in order to develop the optimal formula for the entrapment of Bifidobacterium lactis 300B in Ca-alginate based granules. Laminar flow drip casting with Brace-Encapsulator was used in order to prepare the granules. The results showed that alginate/pullulan and alginate/HPMC fo...

  1. Lactobacillus casei Zhang和Bifidobacterium lactis V9在益生菌酸乳中的应用%Application of Lactobacillus casei Zhang and Bifidobacterium lactis V9 in the probiotic yoghurt

    Institute of Scientific and Technical Information of China (English)

    刘彪; 张和平

    2012-01-01

    益生菌在酸乳中的应用已非常普遍,将Lactobacillus casei Zhang单独(样品A)以及与Bifidobacterium lactis V9复合(样品B),同酸乳发酵剂(G027)共同发酵益生菌酸乳,于4℃贮藏21 d.结果表明,整个贮藏期间2组样品间的黏度和持水性差异不显著;贮藏期间2组样品间L.casei Zhang的活菌数没有差异,且L.casei Zhang和B.lactis V9的活菌数不随贮藏时间而降低;L.casei Zhang和B.lactis V9复合益生菌酸奶感官评价优于单独添加L.casei Zhang酸乳.L.casei Zhang和B.lactis V9复合添加,更适合于益生菌酸乳的生产.%Now it is common for adding probiotics in fermented yoghurt. Yoghurt was fermented by Lactobadllus casei Zhang (A) or combined with Bifidobacterium lactis V9 (B) plus with starter culture (G027) and followed with a 21 d storage at 4 t. The results showed that there was no significance between A and B concerning viscosity and syneresis. It also observed no significant difference existed in counts of L. casei 7ian% between A and B during storage and counts of two probiotics had no change as the storage passed by. However, Fermented milk by combined L. casei Zhang with B. lactis V9 possess markedly higher favorable sensory scores than L. casei Zhang fermented milk, suggesting that it is more suitable for probiotic yoghurt manufacture.

  2. 荧光定量PCR法检测益生菌饮料中Lactobacillus casei Zhang和Bifidobacterium lactis V9%Lactobacillus casei Zhang Bifidobacterium lactis V9 Method of fluorescent quantitative PCR for detection of Lactobacillus casei Zhang and Bifidobacterium lactis V9

    Institute of Scientific and Technical Information of China (English)

    其木格苏都; 王记成; 张家超; 张和平

    2011-01-01

    益生菌活菌数是益生菌产品最重要的指标,而检测益生菌方法的准确性和科学性则至关重要.本文采用荧光定量PCR法同时定量检测益生菌饮料中Lactobacillus casei Zhang(L.casei Zhang)和Bifidobacterium lactis V9(B.lactis V9)的活菌数,并与平板菌落计数法进行比较.结果表明,荧光定量PCR法测得L.casei Zhang活菌数与平板菌落计数法测得活菌数差异不显著;而采用荧光定量PCR法测得B.lactis V9活菌数显著高于平板菌落计数法.荧光定量PCR法灵敏、特异、简便快速,可定量检测并真实反应L.casei Zhang和B.lactis V9的活菌数.%The viable count of probiotic is considered as the most important index and the accurate and scientific assay of probiotic viable number is essential to probiotic products. In the present study, the fluorescent quantitative PCR method and plate colony counting method were used and compared for detection of bacterial viable numbers of Lactobadllus casei Zhang and Bifidobacterium lactis V9 in probiotic fermented beverage. The result showed that there was no significant difference of L. Casei Zhang number between two methods, whereas the numbers of B. Lactis V9 detected by two methods were significant difference. It is suggested that fluorescent quantitative PCR method appear to be highly accurate, specific, fast and reliable for quantification of L. Casei Zhang and B. Lactis V9.

  3. The extracellular proteome of Bifidobacterium animalis subsp. lactis BB‐12 reveals proteins with putative roles in probiotic effects

    DEFF Research Database (Denmark)

    Gilad, Ofir; Svensson, Birte; Viborg, Alexander Holm

    2011-01-01

    Probiotics are live microorganisms that exert health‐promoting effects on the human host, as demonstrated for numerous strains of the genus Bifidobacterium. To unravel the proteins involved in the interactions between the host and the extensively used and well‐studied probiotic strain Bifidobacte......Probiotics are live microorganisms that exert health‐promoting effects on the human host, as demonstrated for numerous strains of the genus Bifidobacterium. To unravel the proteins involved in the interactions between the host and the extensively used and well‐studied probiotic strain...... Bifidobacterium animalis subsp. lactis BB‐12, proteins secreted by the bacterium, i.e. belonging to the extracellular proteome present in the culture medium, were identified by 2‐DE coupled with MALDI‐TOF MS. Among the 74 distinct proteins identified, 31 are predicted to carry out their physiological role either...

  4. The Science behind the Probiotic Strain Bifidobacterium animalis subsp. lactis BB-12®

    Directory of Open Access Journals (Sweden)

    Mikkel Jungersen

    2014-03-01

    Full Text Available This review presents selected data on the probiotic strain Bifidobacterium animalis subsp. lactis BB-12® (BB-12®, which is the world’s most documented probiotic Bifidobacterium. It is described in more than 300 scientific publications out of which more than 130 are publications of human clinical studies. The complete genome sequence of BB-12® has been determined and published. BB-12® originates from Chr. Hansen’s collection of dairy cultures and has high stability in foods and as freeze dried powders. Strain characteristics and mechanisms of BB-12® have been established through extensive in vitro testing. BB-12® exhibits excellent gastric acid and bile tolerance; it contains bile salt hydrolase, and has strong mucus adherence properties, all valuable probiotic characteristics. Pathogen inhibition, barrier function enhancement, and immune interactions are mechanisms that all have been demonstrated for BB-12®. BB-12® has proven its beneficial health effect in numerous clinical studies within gastrointestinal health and immune function. Clinical studies have demonstrated survival of BB-12® through the gastrointestinal tract and BB-12® has been shown to support a healthy gastrointestinal microbiota. Furthermore, BB-12® has been shown to improve bowel function, to have a protective effect against diarrhea, and to reduce side effects of antibiotic treatment, such as antibiotic-associated diarrhea. In terms of immune function, clinical studies have shown that BB-12® increases the body’s resistance to common respiratory infections as well as reduces the incidence of acute respiratory tract infections.

  5. Studies on the storage stability of Bifidobacterium lactis V9 in skim milk powder%Bifidobacterium lactis V9在脱脂乳粉中贮藏稳定性的研究

    Institute of Scientific and Technical Information of China (English)

    刘彪; 张和平

    2012-01-01

    To investigate the bacterial survive stability (B. lactis V9 and Bifidobacterium B) in skim milk powder under different temperatures. Arrhenius equation was involved for evaluating the relationship between temperature and loss rate of bacterial cells in order to predict the loss of B. lactis V9 and Bifidobacterium B in skim milk powder during shelf life. The results showed drat loss rate of bacterial cells was increased as the temperature increment. After at 37 ℃ for 28 days storage, living numbers of two strains were the mosdy reduced by two order of magnitude. No significant changes of living numbers with rime variation was observed at each temperature. A prediction model of shelf life with two strains under different temperatures was established according to Arrhenius equation and bacterial decrement data.This model can predict the loss rate of bacterial cells under one temperature and shelf life of bifidobacteria skim milk powder.%研究Bifidobacterium laais V9(B.laais V9)与双歧杆菌B在脱脂乳粉中不同温度条件下存活稳定性,并以Arrhenius方程反映温度与菌体损失率之间关系,预测B.lactis V9与双歧杆菌B相关乳粉产品货架期内菌体损失情况.结果表明,B.lactis V9与双歧杆菌B随温度升高,菌体损失率增大.两菌株在37℃贮藏28 d后,活菌数下降最明显,约两个数量级,且在不同贮藏温度下活菌数随时间变化情况无显著性差异.通过Arrhenius方程和菌体递减模型,建立了B.lactis V9和双歧杆菌B在不同温度下货架期预测模型,该模型可以较好的预测含B.lactis V9和双歧杆菌B乳粉产品在某一温度下菌体损失率和货架期寿命.

  6. Predominant genera of fecal microbiota in children with atopic dermatitis are not altered by intake of probiotic bacteria Lactobacillus acidophilus NCFM and Bifidobacterium animalis subsp. lactis Bi-07.

    Science.gov (United States)

    Larsen, Nadja; Vogensen, Finn K; Gøbel, Rikke; Michaelsen, Kim F; Abu Al-Soud, Waleed; Sørensen, Søren J; Hansen, Lars H; Jakobsen, Mogens

    2011-03-01

    The effect of probiotic bacteria Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07 on the composition of the Lactobacillus group, Bifidobacterium and the total bacterial population in feces from young children with atopic dermatitis was investigated. The study included 50 children randomized to intake of one of the probiotic strain or placebo. Microbial composition was characterized by denaturing gradient gel electrophoresis, quantitative PCR and, in a subset of subjects, by pyrosequencing of the 16S rRNA gene. The core population of the Lactobacillus group was identified as Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus oris, Leuconostoc mesenteroides, while the bifidobacterial community included Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium longum and Bifidobacterium catenulatum. The fecal numbers of L. acidophilus and B. lactis increased significantly after intervention, indicating survival of the ingested bacteria. The levels of Bifidobacterium correlated positively (P=0.03), while the levels of the Lactobacillus group negatively (P=0.01) with improvement of atopic eczema evaluated by the Severity Scoring of Atopic Dermatitis index. This correlation was observed across the whole study cohort and not attributed to the probiotic intake. The main conclusion of the study is that administration of L. acidophilus NCFM and B. lactis Bi-07 does not affect the composition and diversity of the main bacterial populations in feces.

  7. Influence of probiotic strain Bifidobacterium animalis subsp. lactis lafti® b94, inulin and transglutaminase on the properties of set- style yoghurt

    Directory of Open Access Journals (Sweden)

    Maja Benković

    2008-05-01

    Full Text Available The aim of this research was to examine the influence of probiotic strain Bifidobacterium animalis subsp. lactis LAFTI® B94, inulin and transglutaminase on quality and sensory properties of set-style yoghurt. Fresh, homogenized milk with 3,3% of milk fat was used for yoghurt production, with addition of Bifidobacterium animalis subsp. lactis LAFTI® B94, inulin and transglutaminase activated during 1h 30 min at 55 °C. Enzyme inactivation was carried out by pasteurization of milk during 15 minutes at 85 °C. Control samples were prepared without addition of probiotic culture, inulin and transglutaminase. Physico-chemical parameters and sensory properties of produced set-style yoghurt have been determined. For reliable identification of probiotic strain Bifidobacterium animalis subsp. lactis LAFTI® B94, isolated from the produced yoghurt, SDS-PAGE of whole cell proteins and PCR with species specific primers for Bifidobacterium were carried out. It has been shown that produced set-style yoghurt with probiotic strain Bifidobacterium animalis subsp. lactis LAFTI® B94, inulin and transglutaminase had higher firmness, less syneresis and better sensory properties than control yoghurt samples. After 28 days of storage the viable count of Bifidobacterium animalis subsp. lactis LAFTI® B94 was higher in samples containing inulin as prebiotic. Fermentation of yoghurt samples containing inulin and transglutaminase lasted shorter than fermentation of samples without inulin and transglutaminase. The presence of high number of probiotic culture (more than 106 cells/mL in produced set yoghurts was confirmed by SDS-PAGE of whole cell proteins and PCR with species specific primers for Bifidobacterium

  8. Dose-response study of probiotic bacteria Bifidobacterium animalis subsp lactis BB-12 and Lactobacillus paracasei subsp paracasei CRL-341 in healthy young adults

    DEFF Research Database (Denmark)

    Larsen, C.N.; Nielsen, S.; Kaestel, P.

    2006-01-01

    Objective: This study was performed to investigate the dose-response effects of supplementation with Bifidobacterium animalis subsp lactis (BB-12) and Lactobacillus paracasei subsp paracasei (CRL-431) on blood lipids, recovery from feces and bowel habits. Changes of the fecal microflora was analy......Objective: This study was performed to investigate the dose-response effects of supplementation with Bifidobacterium animalis subsp lactis (BB-12) and Lactobacillus paracasei subsp paracasei (CRL-431) on blood lipids, recovery from feces and bowel habits. Changes of the fecal microflora...

  9. Combined Transcriptome and Proteome Analysis of Bifidobacterium animalis subsp. lactis BB-12 Grown on Xylo-Oligosaccharides and a Model of Their Utilization

    DEFF Research Database (Denmark)

    Gilad, Ofir; Jacobsen, Susanne; Stuer-Lauridsen, B.

    2010-01-01

    Recent studies have demonstrated that xylo-oligosaccharides (XOS), which are classified as emerging prebiotics, selectively enhance the growth of bifidobacteria in general and of Bifidobacterium animalis subsp. lactis strains in particular. To elucidate the metabolism of XOS in the well-documente......Recent studies have demonstrated that xylo-oligosaccharides (XOS), which are classified as emerging prebiotics, selectively enhance the growth of bifidobacteria in general and of Bifidobacterium animalis subsp. lactis strains in particular. To elucidate the metabolism of XOS in the well...

  10. Bifidobacterium animalis subsp. lactis BB-12 in reducing the risk of infections in infancy.

    Science.gov (United States)

    Taipale, Teemu; Pienihäkkinen, Kaisu; Isolauri, Erika; Larsen, Charlotte; Brockmann, Elke; Alanen, Pentti; Jokela, Jorma; Söderling, Eva

    2011-02-01

    The impact of controlled administration of Bifidobacterium animalis subsp. lactis BB-12 (BB-12) on the risk of acute infectious diseases was studied in healthy newborn infants. In this double-blind, placebo-controlled study, 109 newborn 1-month-old infants were assigned randomly to a probiotic group receiving a BB-12-containing tablet (n 55) or to a control group receiving a control tablet (n 54). Test tablets were administered to the infants twice a day (daily dose of BB-12 10 billion colony-forming units) from the age of 1-2 months to 8 months with a novel slow-release pacifier or a spoon. Breastfeeding habits, pacifier use, dietary habits, medications and all signs and symptoms of acute infections were registered. At the age of 8 months, faecal samples were collected for BB-12 determination (quantitative PCR method). The baseline characteristics of the two groups were similar, as was the duration of exclusive breastfeeding. BB-12 was recovered (detection limit log 5) in the faeces of 62% of the infants receiving the BB-12 tablet. The daily duration of pacifier sucking was not associated with the occurrence of acute otitis media. No significant differences between the groups were observed in reported gastrointestinal symptoms, otitis media or use of antibiotics. However, the infants receiving BB-12 were reported to have experienced fewer respiratory infections (65 v. 94%; risk ratio 0·69; 95% CI 0·53, 0·89; P = 0·014) than the control infants. Controlled administration of BB-12 in early childhood may reduce respiratory infections.

  11. Resistance determinant erm(X) is borne by transposon Tn5432 in Bifidobacterium thermophilum and Bifidobacterium animals subsp. lactis

    NARCIS (Netherlands)

    Hoek, van A.H.A.M.; Mayrhofer, S.; Domig, K.J.; Aarts, H.J.M.

    2008-01-01

    The erm(X) gene from erythromycin- and clindamycin-resistant Bifidobacterium strains was characterised by polymerase chain reaction and sequence analysis, including flanking regions. Results suggest that the resistance determinant was part of transposon Tn5432 that has been described in several oppo

  12. Inhibitory effect of honey-sweetened goat and cow milk fermented with Bifidobacterium lactis Bb-12 on the growth of Listeria monocytogenes

    Directory of Open Access Journals (Sweden)

    Mirela Lučan

    2009-06-01

    Full Text Available The aim of the study was to determine the influence of honey addition on fermentation of goat and cow milk with Bifidobacterium lactis Bb-12. Additionally, inhibitory potential of honey-sweetened fermented goat and cow milk against Listeria monocytogenes strain was examined. Two monofloral honey types, dark-colored chestnut and light-colored acacia honey were added. The basic hypothesis of this study was that addition of honey could have influence on the growth of Bifidobacterium lactis during the fermentation of goat and cow milk. Furthermore, higher inhibitory potential caused by honey addition against Listeria monocytogenes has been assumed. Compared to cow milk, higher acidity and CFU of Bifidobacterium lactis Bb-12 were noted in the fermented goat milk in all phases of the fermentation process. The results of this study show that both types of honey enhanced growth and acidity of the Bifidobacterium lactis Bb-12 in both milk types during fermentation. A disc assay has shown that development of growth inhibition zones depends on the type and concentration of honey, as well as on the milk type. The chestnut honey had generally higher inhibitory effect than acacia honey.

  13. Effect of a fermented milk containing Bifidobacterium lactis DN-173010 on Chinese constipated women

    Institute of Scientific and Technical Information of China (English)

    Yue-Xin Yang; Mei He; Gang Hu; Jie Wei; Philippe Pages; Xian-Hua Yang; Sophie Bourdu-Naturel

    2008-01-01

    AIM: To investigate the effect of a fermented milk containing Bifidobacterium lactis DN- 173010 and yogurt strains (BIO ) on adult women with constipation in Beijing. METHODS: A total of 135 adult females with constipation were randomly allocated to consume for 2 wk either 100 g of the test fermented milk or 100 g of an acidified milk containing non-living bacteria (control). Stool frequency, defecation condition scores, stool consistency and food intake were recorded at baseline and after 1 and 2 wk in an intention-to-treat population of 126 subjects. In parallel, safety evaluation parameters were performed. RESULTS: At baseline, no differences were found between groups. Following consumption of test product, stool frequency was significantly increased after 1 wk (3.5±1.5 vs 2.4±0.6, P < 0.01) and 2 wk (4.1±1.7 vs 2.4±0.6, P < 0.01), vs baseline. Similarly, after 1 and 2 wk, of test product consumption, defecation condition (1.1±0.9 vs 1.9±1.2, P < 0.01 and 0.8±1.0 vs 1.9±1.2, P < 0.01, respectively) and stool consistency (1.0±0.8 vs 1.5±1.1, P < 0.01 and 0.6±0.8 vs 1.5±1.1, P < 0.01, respectively) were significantly improved. Compared with the control group, stool frequency was also significantly increased (3.5±1.5 vs 2.5± 0.9, P <0.01 and 4.1±1.7 vs 2.6±1.0, P <0.01, respectively), and defecation condition (1.1±0.9 vs 1.6±1.1, P < 0.01 and 0.8±1.0 vs 1.6±1.1, P < 0.01, respectively) and stool consistency (1.0±0.8 vs 1.4±1.0, P < 0.05 and 0.6±0.8 vs 1.3±1.0, P < 0.01, respectively) significantly decreased after 1 and 2 wk of product consumption. During the same period, food intake did not change between the two groups, and safety parameters of the subjects were within normal ranges. CONCLUSION: This study suggests a beneficial effect of a fermented milk containing B. lactis DN-173010 on stool frequency, defecation condition and stool consistency in adult women with constipation constipated women after 1 and 2 wk of consumption.

  14. Predominant genera of fecal microbiota in children with atopic dermatitis are not altered by intake of probiotic bacteria Lactobacillus acidophilus NCFM and Bifidobacterium animalis subsp.lactis Bi-07

    DEFF Research Database (Denmark)

    Larsen, Nadejda Nikolajevna; Vogensen, Finn Kvist; Gøbel, Rikke Juul

    2011-01-01

    The effect of probiotic bacteria Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07 on the composition of the Lactobacillus group, Bifidobacterium and the total bacterial population in feces from young children with atopic dermatitis was investigated. The study included 50 children...... as Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus oris, Leuconostoc mesenteroides, while the bifidobacterial community included Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium longum and Bifidobacterium catenulatum. The fecal numbers of L. acidophilus and B. lactis increased...... significantly after intervention, indicating survival of the ingested bacteria. The levels of Bifidobacterium correlated positively (P = 0.03), while the levels of the Lactobacillus group negatively (P = 0.01) with improvement of atopic eczema evaluated by the Severity Scoring of Atopic Dermatitis index...

  15. Safety of Bifidobacterium animalis subsp. lactis (B. lactis) strain BB-12-supplemented yogurt in healthy adults on antibiotics: a phase I safety study.

    Science.gov (United States)

    Merenstein, Daniel J; Tan, Tina P; Molokin, Aleksey; Smith, Keisha Herbin; Roberts, Robert F; Shara, Nawar M; Mete, Mihriye; Sanders, Mary Ellen; Solano-Aguilar, Gloria

    2015-01-01

    Probiotics are live microorganisms that, when administered in sufficient doses, provide health benefits on the host. The United States Food and Drug Administration (FDA) requires phase I safety studies for probiotics when the intended use of the product is as a drug. The purpose of the study was to determine the safety of Bifidobacterium animalis subsp lactis (B. lactis) strain BB-12 (BB-12)-supplemented yogurt when consumed by a generally healthy group of adults who were prescribed a 10-day course of antibiotics for a respiratory infection. Secondary aims were to assess the ability of BB-12 to affect the expression of whole blood immune markers associated with cell activation and inflammatory response. A phase I, double-blinded, randomized controlled study was conducted in compliance with FDA guidelines for an Investigational New Drug (IND). Forty participants were randomly assigned to consume 4 ounces of either BB-12 -supplemented yogurt or non-supplemented control yogurt daily for 10 d. The primary outcome was to assess safety and tolerability, assessed by the number of reported adverse events. A total of 165 non-serious adverse events were reported, with no differences between the control and BB-12 groups. When compared to the control group, B lactis fecal levels were modestly higher in the BB-12-supplemented group. In a small subset of patients, changes in whole blood expression of genes associated with regulation and activation of immune cells were detected in the BB-12-supplemented group. BB-12-supplemented yogurt is safe and well tolerated when consumed by healthy adults concurrently taking antibiotics. This study will form the basis for future randomized clinical trials investigating the potential immunomodulatory effects of BB-12-supplemented yogurt in a variety of disease states.

  16. The influence of different polymers on viability of Bifidobacterium lactis 300b during encapsulation, freeze-drying and storage.

    Science.gov (United States)

    Pop, Oana Lelia; Brandau, Thorsten; Schwinn, Jens; Vodnar, Dan Cristian; Socaciu, Carmen

    2015-07-01

    Seven different types of natural polymers namely hydroxypropyl methylcellulose (HPMC), sodium-carboxymethyl cellulose (Na-CMC), microcrystalline cellulose (MCC), starch BR-07, starch BR-08, dextrin and pullulan were used in order to develop the optimal formula for the entrapment of Bifidobacterium lactis 300B in Ca-alginate based granules. Laminar flow drip casting with Brace-Encapsulator was used in order to prepare the granules. The results showed that alginate/pullulan and alginate/HPMC formulation provide high protection for the bacterial strain used for encapsulation. These two formulations were further used to obtain freeze dried granules, for which the viability in time and at different temperatures was tested. The final results showed a higher viability than the level of the therapeutic minimum (>10(7) CFU/g) after 15 days of storage. Other parameters like entrapment efficiency, production rate, sphericity, flowability were also discussed.

  17. Probiotic Lactobacillus acidophilus NCFM and Bifidobacterium animalis subsp lactis Bl-04 interactions with prebiotic carbohydrates using differential proteomics and protein characterization

    DEFF Research Database (Denmark)

    Hansen, Morten Ejby

    are defined as probiotics. The positive clinical effects of probiotics, mainly belonging to the Bifidobacterium and Lactobacillus genera in treatments of irritable bowel syndrome, gut infections and lifestyle diseases are well documented. Compounds that selectively stimulate the beneficial effect...... and the discovery and documentation of novel ones. In this project we set out to investigate the metabolism of carbohydrates that are prebiotic or potential prebiotic compounds utilized by the probiotic organisms Lactobacillus acidophilus NCFM (NCFM) and Bifidobacterium animalis subsp. lactis BL-04 (Bl-04). The aim...

  18. Mortality and translocation assay to study the protective capacity of Bifidobacterium lactis INL1 against Salmonella Typhimurium infection in mice.

    Science.gov (United States)

    Zacarías, M F; Reinheimer, J; Forzani, L; Grangette, C; Vinderola, G

    2014-12-01

    The mouse has been largely used for the study of the protective capacity of probiotics against intestinal infections caused by Salmonella. In this work we aimed at comparing the mortality and translocation assay for the study of the protective capacity of the human breast milk-derived strain Bifidobacterium animalis subsp. lactis INL1 on a model of gut infection by Salmonella enterica subsp. enterica serovar Typhimurium. Different doses of S. Typhimurium FUNED and B. animalis subsp. lactis INL 1 were administered to Balb/c mice in a mortality or a translocation assay. The survival of the control group in the mortality assay resulted to be variable along experiments, and then we preferred to use a translocation assay where the preventive administration of 109 cfu of bifidobacteria/mouse for 10 consecutive days significantly reduced the number of infected animals and the levels of translocation to liver and spleen, with enhanced secretory immunoglobulin A and interleukin 10 production in the small and large intestine, respectively. Ten days of B. animalis subsp. lactis strain INL1 administration to mice significantly reduced both the incidence and the severity of Salmonella infection in a mouse model of translocation. This work provided the first evidence that a translocation assay, compared to a mortality assay, could be more useful to study the protective capacity of probiotics against Salmonella infection, as more information can be obtained from mice and less suffering is conferred to animals due to the fact that the mortality assay is shorter than the latter. These facts are in line with the guidelines of animal research recently established by the National Centre for the Replacement, Refinement & Reduction of Animals in Research.

  19. 不同气体环境对益生菌Bifidobacterium lactis V9生长的影响%Effects of air condition on the viability of probiotic bacteria: Bifidobacterium lactis V9

    Institute of Scientific and Technical Information of China (English)

    其木格苏都; 白梅; 孔亚楠; 魏爱彬; 王记成; 张和平

    2012-01-01

    Bifidobacterium lactis V9(B.lactis V9)是一株具有良好益生特性且遗传稳定的益生茵,工业化生产环境中气体组成关系着益生菌活菌数量,进而影响其益生功效.[目的]研究不同气体环境对B.lactis V9生长及代谢的影响.[方法]在固体MRS培养基、液体MRS培养基及巴氏杀菌脱脂乳接种B.lactis V9,于不同的气体环境中培养.[结果]在固体MRS培养基上,B.lactis V9在混合气体(N2∶H2∶CO2=80∶10∶10)环境中菌落形成较氮气环境(N2∶99.99%)多,在空气环境(N2∶O2≈79∶21)中菌落形成极少.B.lactis V9在MRS液体中培养24 h,混合气体环境下其活菌数(9.11±0.11 log CFU/mL)显著高于空气环境下的活菌数(8.04±0.10 log CFU/mL) (P<0.01),在混合气体环境下B.lactis V9代谢生成的乙酸和乳酸量分别为12.79±0.86 mmol/L和11.99±0.73 mmol/L,显著高于在空气环境中生成量0.65±0.07 mmol/L和2.75±0.57 mmol/L (P<0.01),乙酸/乳酸比值分别为1.06∶1和0.24∶1.B.lactis V9在巴氏杀菌脱脂乳中发酵18h,混合气体环境下pH值(4.48±0.07)显著低于空气环境下的pH值(5.03±0.12) (P<0.01),混合气体环境下其活菌数(9.02±0.15 log CFU/mL)显著高于空气环境下的活菌数(8.53±0.08 log CFU/mL) (P<0.01).混合气体和空气环境下发酵脱脂乳产生的乙酸和乳酸量分别为60.52±2.30 mmol/L、5.17±1.02 mmol/L和16.86±0.34 mmol/L、5.92±0.81 mmol/L,乙酸/乳酸的值分别为11.71∶1和2.85∶1.[结论]在N2∶H2∶CO2=80∶10∶10混合气体环境下有利于B.lactis V9在液体MRS和脱脂乳中生长,其活菌数可以增加0.5-1个数量级.这一研究结果也可为B.lactis V9益生菌发酵乳的生产和产品中B.lactis V9活菌培养计数提供指导.%Bifldobacterium lactis V9 (B.lactis V9) has been demostrated as a probiotic with well properties and stable genetics.The gas composition in environment of industrial manufacturation is positively associated with viable numbers of

  20. Probiotic treatment of collagenous colitis: a randomized, double-blind, placebo-controlled trial with Lactobacillus acidophilus and Bifidobacterium animalis subsp. Lactis

    DEFF Research Database (Denmark)

    Wildt, Signe; Munck, Lars K; Vinter-Jensen, Lars

    2006-01-01

    Probiotic treatment may be effective in diseases involving gut microflora and intestinal inflammation. In collagenous colitis (CC), a potential pathogenic role of the gut microflora has been proposed. The effect of probiotic treatment in CC is unknown. Our aim was to investigate the clinical effe...... of treatment with Lactobacillus acidophilus LA-5 and Bifidobacterium animalis subsp. lactis BB-12 (AB-Cap-10) in patients with CC....

  1. Discovery of proteins involved in the interaction between prebiotics carbohydrates and probiotics & whole proteome analysis of the probiotic strain Bifidobacterium animalis susp. lactis BB-12

    DEFF Research Database (Denmark)

    Gilad, Ofir

    Probiotic bacteria, which primarily belong to the genera Lactobascillus and Bifidobacterium, are live microorganisms that have been related to a variety of health-promoting effects. Prebiotics are indigestible food components that specifically stimulate the growth of probiotic organisms in the hu......Probiotic bacteria, which primarily belong to the genera Lactobascillus and Bifidobacterium, are live microorganisms that have been related to a variety of health-promoting effects. Prebiotics are indigestible food components that specifically stimulate the growth of probiotic organisms...... between the widely-used, extensively-studied probiotic strain Bifidobacterium animalis subsp. lactis BB-12 and potentially-prebiotic carbohydrates. The project was initiated with a screening phase in which more than 40 carbohydrates were tested for their ability to promote the growth of the bacterium...

  2. Effect of probiotic yogurt containing Lactobacillus acidophilus and Bifidobacterium lactis on lipid profile in individuals with type 2 diabetes mellitus.

    Science.gov (United States)

    Ejtahed, H S; Mohtadi-Nia, J; Homayouni-Rad, A; Niafar, M; Asghari-Jafarabadi, M; Mofid, V; Akbarian-Moghari, A

    2011-07-01

    The purpose of this study was to investigate the effects of probiotic and conventional yogurt on the lipid profile in type 2 diabetic people. In a randomized double-blind controlled trial, 60 people (23 males and 37 females) with type 2 diabetes and low-density lipoprotein cholesterol (LDL-C) greater than 2.6 mmol/L were assigned to 2 groups. Participants consumed daily 300 g of probiotic yogurt containing Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 or 300 g of conventional yogurt for 6 wk. Fasting blood samples, anthropometric measurements and 3-d, 24-h dietary recalls were collected at the baseline and at the end of the trial. Probiotic yogurt consumption caused a 4.54% decrease in total cholesterol and a 7.45% decrease in LDL-C compared with the control group. No significant changes from baseline were shown in triglyceride and high-density lipoprotein cholesterol (HDL-C) in the probiotic group. The total cholesterol:HDL-C ratio and LDL-C:HDL-C ratio as atherogenic indices significantly decreased in the probiotic group compared with the control group. Probiotic yogurt improved total cholesterol and LDL-C concentrations in type 2 diabetic people and may contribute to the improvement of cardiovascular disease risk factors.

  3. Catabolism of glucose and lactose in Bifidobacterium animalis subsp. lactis, studied by 13C Nuclear Magnetic Resonance.

    Science.gov (United States)

    González-Rodríguez, Irene; Gaspar, Paula; Sánchez, Borja; Gueimonde, Miguel; Margolles, Abelardo; Neves, Ana Rute

    2013-12-01

    Bifidobacteria are widely used as probiotics in several commercial products; however, to date there is little knowledge about their carbohydrate metabolic pathways. In this work, we studied the metabolism of glucose and lactose in the widely used probiotic strain Bifidobacterium animalis subsp. lactis BB-12 by in vivo (13)C nuclear magnetic resonance (NMR) spectroscopy. The metabolism of [1-(13)C]glucose was characterized in cells grown in glucose as the sole carbon source. Moreover, the metabolism of lactose specifically labeled with (13)C on carbon 1 of the glucose or the galactose moiety was determined in suspensions of cells grown in lactose. These experiments allowed the quantification of some intermediate and end products of the metabolic pathways, as well as determination of the consumption rate of carbon sources. Additionally, the labeling patterns in metabolites derived from the metabolism of glucose specifically labeled with (13)C on carbon 1, 2, or 3 in cells grown in glucose or lactose specifically labeled in carbon 1 of the glucose moiety ([1-(13)Cglucose]lactose), lactose specifically labeled in carbon 1 of the galactose moiety ([1-(13)Cgalactose]lactose), and [1-(13)C]glucose in lactose-grown cells were determined in cell extracts by (13)C NMR. The NMR analysis showed that the recovery of carbon was fully compatible with the fructose 6-phosphate, or bifid, shunt. The activity of lactate dehydrogenase, acetate kinase, fructose 6-phosphate phosphoketolase, and pyruvate formate lyase differed significantly between glucose and lactose cultures. The transcriptional analysis of several putative glucose and lactose transporters showed a significant induction of Balat_0475 in the presence of lactose, suggesting a role for this protein as a lactose permease. This report provides the first in vivo experimental evidence of the metabolic flux distribution in the catabolic pathway of glucose and lactose in bifidobacteria and shows that the bifid shunt is the only

  4. Biochemical and kinetic characterisation of a novel xylooligosaccharide-upregulated GH43 β-d-xylosidase/α-l-arabinofuranosidase (BXA43) from the probiotic Bifidobacterium animalis subsp. lactis BB-12

    DEFF Research Database (Denmark)

    Viborg, Alexander Holm; Sørensen, Kim Ib; Gilad, Ofir

    2013-01-01

    The Bifidobacterium animalis subsp. lactis BB-12 gene BIF_00092, assigned to encode a β-d-xylosidase (BXA43) of glycoside hydrolase family 43 (GH43), was cloned with a C-terminal His-tag and expressed in Escherichia coli. BXA43 was purified to homogeneity from the cell lysate and found to be a dual...... to xylotetraose at subsite +3 and occur only in GH43 from the Bifidobacterium genus....

  5. Preservation of functionality of Bifidobacterium animalis subsp. lactis INL1 after incorporation of freeze-dried cells into different food matrices.

    Science.gov (United States)

    Vinderola, G; Zacarías, M F; Bockelmann, W; Neve, H; Reinheimer, J; Heller, K J

    2012-05-01

    The aim of this work was to investigate how production and freeze-drying conditions of Bifidobacterium animalis subsp. lactis INL1, a probiotic strain isolated from breast milk, affected its survival and resistance to simulated gastric digestion during storage in food matrices. The determination of the resistance of bifidobacteria to simulated gastric digestion was useful for unveiling differences in cell sensitivity to varying conditions during biomass production, freeze-drying and incorporation of the strain into food products. These findings show that bifidobacteria can become sensitive to technological variables (biomass production, freeze-drying and the food matrix) without this fact being evidenced by plate counts.

  6. Effect of yoghurt containing Bifidobacterium lactis Bb12® on faecal excretion of secretory immunoglobulin A and human beta-defensin 2 in healthy adult volunteers

    Directory of Open Access Journals (Sweden)

    Kabeerdoss Jayakanthan

    2011-12-01

    Full Text Available Abstract Background Probiotics are used to provide health benefits. The present study tested the effect of a probiotic yoghurt on faecal output of beta-defensin and immunoglobulin A in a group of young healthy women eating a defined diet. Findings 26 women aged 18-21 (median 19 years residing in a hostel were given 200 ml normal yoghurt every day for a week, followed by probiotic yoghurt containing Bifidobacterium lactis Bb12® (109 in 200 ml for three weeks, followed again by normal yoghurt for four weeks. Stool samples were collected at 0, 4 and 8 weeks and assayed for immunoglobulin A and human beta-defensin-2 by ELISA. All participants tolerated both normal and probiotic yoghurt well. Human beta-defensin-2 levels in faeces were not altered during the course of the study. On the other hand, compared to the basal sample, faecal IgA increased during probiotic feeding (P = 0.0184 and returned to normal after cessation of probiotic yoghurt intake. Conclusions Bifidobacterium lactis Bb12® increased secretory IgA output in faeces. This property may explain the ability of probiotics to prevent gastrointestinal and lower respiratory tract infections.

  7. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) Scientific Opinion on the substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and

    DEFF Research Database (Denmark)

    Tetens, Inge

    substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and improves stools frequency. The food that is the subject of the health claim is a combination of B. longum LA 101, L...

  8. Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 cell surface hydrophobicity and survival of the cells under adverse environmental conditions.

    Science.gov (United States)

    Shakirova, Laisana; Grube, Mara; Gavare, Marita; Auzina, Lilija; Zikmanis, Peteris

    2013-01-01

    Changes in the cell surface hydrophobicity (CSH) of probiotic bacteria Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 and the survival of these cells were examined in response to varied cultivation conditions and adverse environmental conditions. An inverse linear relationship (P acidophilus La5 and B. lactis Bb12 and survival of cells subjected to subsequent freezing/thawing, long-term storage or exposure to mineral and bile acids. The observed relationships were supported by significant correlations between the CSH and changes in composition of the cell envelopes (proteins, lipids and carbohydrates) of L. acidophilus La5 and B. lactis Bb12 examined using FT-IR spectroscopy and conventional biochemical analysis methods. The results also suggest that the estimates of hydrophobicity, being a generalized characteristic of cell surfaces, are important parameters to predict the ability of intact probiotic bacteria to endure extreme environments and therefore should be monitored during cultivation. A defined balance of cell components, which can be characterized by the reduced CSH values, apparently helps to ensure the resistance, improved viability and hence the overall probiotic properties of bacteria.

  9. Spray-drying process preserves the protective capacity of a breast milk-derived Bifidobacterium lactis strain on acute and chronic colitis in mice

    Science.gov (United States)

    Burns, Patricia; Alard, Jeanne; Hrdỳ, Jiri; Boutillier, Denise; Páez, Roxana; Reinheimer, Jorge; Pot, Bruno; Vinderola, Gabriel; Grangette, Corinne

    2017-01-01

    Gut microbiota dysbiosis plays a central role in the development and perpetuation of chronic inflammation in inflammatory bowel disease (IBD) and therefore is key target for interventions with high quality and functional probiotics. The local production of stable probiotic formulations at limited cost is considered an advantage as it reduces transportation cost and time, thereby increasing the effective period at the consumer side. In the present study, we compared the anti-inflammatory capacities of the Bifidobacterium animalis subsp. lactis (B. lactis) INL1, a probiotic strain isolated in Argentina from human breast milk, with the commercial strain B. animalis subsp. lactis BB12. The impact of spray-drying, a low-cost alternative of bacterial dehydration, on the functionality of both bifidobacteria was also investigated. We showed for both bacteria that the spray-drying process did not impact on bacterial survival nor on their protective capacities against acute and chronic colitis in mice, opening future perspectives for the use of strain INL1 in populations with IBD. PMID:28233848

  10. Identification of restriction-modification systems of Bifidobacterium animalis subsp. lactis CNCM I-2494 by SMRT sequencing and associated methylome analysis.

    Science.gov (United States)

    O' Connell Motherway, Mary; Watson, Debbie; Bottacini, Francesca; Clark, Tyson A; Roberts, Richard J; Korlach, Jonas; Garault, Peggy; Chervaux, Christian; van Hylckama Vlieg, Johan E T; Smokvina, Tamara; van Sinderen, Douwe

    2014-01-01

    Bifidobacterium animalis subsp. lactis CNCM I-2494 is a component of a commercialized fermented dairy product for which beneficial effects on health has been studied by clinical and preclinical trials. To date little is known about the molecular mechanisms that could explain the beneficial effects that bifidobacteria impart to the host. Restriction-modification (R-M) systems have been identified as key obstacles in the genetic accessibility of bifidobacteria, and circumventing these is a prerequisite to attaining a fundamental understanding of bifidobacterial attributes, including the genes that are responsible for health-promoting properties of this clinically and industrially important group of bacteria. The complete genome sequence of B. animalis subsp. lactis CNCM I-2494 is predicted to harbour the genetic determinants for two type II R-M systems, designated BanLI and BanLII. In order to investigate the functionality and specificity of these two putative R-M systems in B. animalis subsp. lactis CNCM I-2494, we employed PacBio SMRT sequencing with associated methylome analysis. In addition, the contribution of the identified R-M systems to the genetic accessibility of this strain was assessed.

  11. Effect of the consumption of a fermented dairy product containing Bifidobacterium lactis DN-173 010 on constipation in childhood: a multicentre randomised controlled trial (NTRTC: 1571

    Directory of Open Access Journals (Sweden)

    Perrin Catherine

    2009-03-01

    Full Text Available Abstract Background Constipation is a frustrating symptom affecting 3% of children worldwide. Randomised controlled trials show that both polyethylene glycol and lactulose are effective in increasing defecation frequency in children with constipation. However, in 30–50%, these children reported abdominal pain, bloating, flatulence, diarrhoea, nausea and bad taste of the medication. Two recent studies have shown that the fermented dairy product containing Bifidobacterium lactis strain DN-173 010 is effective in increasing stool frequency in constipation-predominant irritable bowel syndrome patients with a defecation frequency Methods/design It is a two nation (The Netherlands and Poland double-blind, placebo-controlled randomised multicentre trial in which 160 constipated children (age 3–16 years with a defecation frequency Bifidobacterium lactis DN-173 010 or a control product, twice a day, for 3 weeks. During the study all children are instructed to try to defecate on the toilet for 5–10 minutes after each meal (3 times a day and daily complete a standardized bowel diary. Primary endpoint is stool frequency. Secondary endpoints are stool consistency, faecal incontinence frequency, pain during defecation, digestive symptoms (abdominal pain, flatulence, adverse effects (nausea, diarrhoea, bad taste and intake of rescue medication (Bisacodyl. Rate of success and rate of responders are also evaluated, with success defined as ≥ 3 bowel movements per week and ≤1 faecal incontinence episode over the last 2 weeks of product consumption and responder defined as a subject reporting a stool frequency ≥ 3 on the last week of product consumption. To demonstrate that the success percentage in the intervention group will be 35% and the success percentage in the control group (acidified milk without ferments, toilet training, bowel diary will be 15%, with alpha 0.05 and power 80%, a total sample size of 160 patients was calculated. Conclusion This

  12. The GH5 1,4-β-mannanase from Bifidobacterium animalis subsp. lactis Bl-04 possesses a low-affinity mannan-binding module and highlights the diversity of mannanolytic enzymes

    DEFF Research Database (Denmark)

    Morrill, Johan; Kulcinskaja, Evelina; Sulewska, Anna Maria

    2015-01-01

    β-Mannans are abundant and diverse plant structural and storage polysaccharides. Certain human gut microbiota members including health-promoting Bifidobacterium spp. catabolize dietary mannans. Little insight is available on the enzymology of mannan deconstruction in the gut ecological niche. Here......, we report the biochemical properties of the first family 5 subfamily 8 glycoside hydrolase (GH5_8) mannanase from the probiotic bacterium Bifidobacterium animalis subsp. lactis Bl-04 (BlMan5_8). BlMan5_8 possesses a novel low affinity carbohydrate binding module (CBM) specific for soluble mannan...

  13. The GH5 1,4-β-mannanase from Bifidobacterium animalis subsp. lactis Bl-04 possesses a low-affinity mannan-binding module and highlights the diversity of mannanolytic enzymes

    DEFF Research Database (Denmark)

    Morrill, Johan; Kulcinskaja, Evelina; Sulewska, Anna Maria;

    2015-01-01

    β-Mannans are abundant and diverse plant structural and storage polysaccharides. Certain human gut microbiota members including health-promoting Bifidobacterium spp. catabolize dietary mannans. Little insight is available on the enzymology of mannan deconstruction in the gut ecological niche. Here......, we report the biochemical properties of the first family 5 subfamily 8 glycoside hydrolase (GH5_8) mannanase from the probiotic bacterium Bifidobacterium animalis subsp. lactis Bl-04 (BlMan5_8). BlMan5_8 possesses a novel low affinity carbohydrate binding module (CBM) specific for soluble mannan...

  14. Development of a rapid SNP-typing assay to differentiate Bifidobacterium animalis ssp. lactis strains used in probiotic-supplemented dairy products.

    Science.gov (United States)

    Lomonaco, Sara; Furumoto, Emily J; Loquasto, Joseph R; Morra, Patrizia; Grassi, Ausilia; Roberts, Robert F

    2015-02-01

    Identification at the genus, species, and strain levels is desirable when a probiotic microorganism is added to foods. Strains of Bifidobacterium animalis ssp. lactis (BAL) are commonly used worldwide in dairy products supplemented with probiotic strains. However, strain discrimination is difficult because of the high degree of genome identity (99.975%) between different genomes of this subspecies. Typing of monomorphic species can be carried out efficiently by targeting informative single nucleotide polymorphisms (SNP). Findings from a previous study analyzing both reference and commercial strains of BAL identified SNP that could be used to discriminate common strains into 8 groups. This paper describes development of a minisequencing assay based on the primer extension reaction (PER) targeting multiple SNP that can allow strain differentiation of BAL. Based on previous data, 6 informative SNP were selected for further testing, and a multiplex preliminary PCR was optimized to amplify the DNA regions containing the selected SNP. Extension primers (EP) annealing immediately adjacent to the selected SNP were developed and tested in simplex and multiplex PER to evaluate their performance. Twenty-five strains belonging to 9 distinct genomic clusters of B. animalis ssp. lactis were selected and analyzed using the developed minisequencing assay, simultaneously targeting the 6 selected SNP. Fragment analysis was subsequently carried out in duplicate and demonstrated that the assay yielded 8 specific profiles separating the most commonly used commercial strains. This novel multiplex PER approach provides a simple, rapid, flexible SNP-based subtyping method for proper characterization and identification of commercial probiotic strains of BAL from fermented dairy products. To assess the usefulness of this method, DNA was extracted from yogurt manufactured with and without the addition of B. animalis ssp. lactis BB-12. Extracted DNA was then subjected to the minisequencing

  15. Fecal water genotoxicity is predictive of tumor-preventive activities by inulin-like oligofructoses, probiotics (Lactobacillus rhamnosus and Bifidobacterium lactis), and their synbiotic combination.

    Science.gov (United States)

    Klinder, Annett; Förster, Antje; Caderni, Giovanna; Femia, Angelo Pietro; Pool-Zobel, Beatrice L

    2004-01-01

    The measurement of fecal water genotoxicity in human colon cells could be a useful biomarker to study effects of diet in the colon. Here we assessed aqueous fecal extracts of samples from a chronic study with rats fed prebiotics, probiotics, and their combination. Treatments were maltodextrins (controls), inulin/oligofructoses (prebiotic), Lactobacillus rhamnosus, and Bifidobacterium lactis (probiotics) or both (synbiotic). Azoxymethane (AOM) was administered to initiate tumors. Rat feces were collected at 0 and 10 days and 2, 4, and 8 mo, and cecal contents were collected at 8 mo. Aqueous phases were prepared and tested for genotoxicity in HT29 colon cells using the comet assay. The studied types of intervention reduced fecal and cecal genotoxicity. DNA damage by samples from AOM-treated, tumor-free rats was significantly lower than from tumor-bearing animals, especially after 4 mo of synbiotic and prebiotic interventions. Inulin-based diets reduced exposure to genotoxins in the feces, directly reflecting the reported reduction of tumor incidence in these animals. Evidence is provided for the validity of this measurement as a biomarker of chemoprevention because 1) fecal water genotoxicity reflected genotoxic exposure in the cecum, 2) tumor incidence and fecal genotoxicity were directly related, and 3) these interventions reduced tumor risks by reducing exposure to genotoxins in the gut.

  16. Genome-wide identification of small RNAs in Bifidobacterium animalis subsp. lactis KLDS 2.0603 and their regulation role in the adaption to gastrointestinal environment.

    Directory of Open Access Journals (Sweden)

    De-Quan Zhu

    Full Text Available Bifidobacteria are one of the predominant bacterial species in the human gastrointestinal tract (GIT and play a vital role in the host's health by acting as probiotics. However, how they regulate themselves to adapt to GIT of their host remains unknown.Eighteen bifidobacterial strains were used to analyze their adaptive capacities towards simulated GIT environment. The strain with highest survival rate and adhesion ability was selected for comparative genome as well as transcriptomic analysis.The Bifidobacterium animalis subsp. lactis KLDS 2.0603 strain was demonstrated to have the highest survival rate and adhesion ability in simulated GIT treatments. The comparative genome analysis revealed that the KLDS 2.0603 has most similar whole genome sequence compared with BB-12 strain. Eleven intergenic sRNAs were identified after genomes prediction and transcriptomic analysis of KLDS 2.0603. Transcriptomic analysis also showed that genes (mainly sRNAs targeted genes and sRNAs were differentially expressed in different stress conditions, suggesting that sRNAs might play a crucial role in regulating genes involved in the stress resistance of this strain towards environmental changes.This study first provided deep and comprehensive insights into the regulation of KLDS 2.0603 strain at transcription and post-transcription level towards environmental.

  17. THE EFFECT OF PROBIOTIC FERMENTED MILK THAT INCLUDES BIFIDOBACTERIUM LACTIS CNCM I-2494 ON THE REDUCTION OF GASTROINTESTINAL DISCOMFORT AND SYMPTOMS IN ADULTS: A NARRATIVE REVIEW.

    Science.gov (United States)

    Waitzberg, Dan L; Quilici, Flávio A; Michzputen, Sender; Friche Passos, Maria do Carmo

    2015-08-01

    Objetivo: determinar la eficacia de la leche fermentada con Bifidobacterium lactis CNCM I-2429 en la reducción de el malestar gastrointestinal (GI) en adultos sanos. Métodos: se realizó una búsqueda sistemática en la literatura para identificar estudios que informaron del uso de B. animalis spp. lactis para molestias/ confort GI en adultos sanos. Se identificaron un total de 5.329 registros, de estos se evaluaron 99 artículos de texto completo. Las búsquedas de ensayos adicionales se realizaron utilizando los nombres de los autores de cada estudio identificado y varias bases de datos relevantes. La selección de los estudios se llevó a cabo de acuerdo con las guías de Artículos de Información Preferidos para Revisiones Sistemáticas y Meta-Análisis (PRISMA). Los estudios eran incluidos si eran ensayos randomizados controlados, si los sujetos de estudio eran adultos sanos y si el grupo de intervención recibió B. lactis CNCM I-2494. Se excluyeron los estudios que no eran randomizados, que incluían adultos que no estaban sanos, que incluían el uso de cualquier otra intervención o si comparaban diferentes productos sin un grupo placebo. La calidad metodológica de los estudios se evaluó utilizando la Escala de Calidad de Oxford y la Evaluación Cochrane de ocultamiento. No fue posible un metaanálisis. Resultados: la estrategia de búsqueda identificó dos estudios que incluyeron un total de 538 mujeres sanas, con edades entre 18 a 60 años, de peso normal o sobrepeso (IMC de 18-30 kg/m2). En uno de los estudios las molestias GI disminuyeron significativamente en el grupo de probióticos frente al grupo control, sin diferencias en el otro. El porcentaje de respondedores para el bienestar GI fue mayor en el grupo de probióticos frente al grupo control en el primer estudio, pero no en el segundo. Los síntomas GI se redujeron significativamente en el grupo probiótico frente al grupo control en ambos estudios. La función intestinal solo se evaluó en

  18. Effect of Lactobacillus rhamnosus LGG® and Bifidobacterium animalis ssp. lactis BB-12® on health-related quality of life in college students affected by upper respiratory infections.

    Science.gov (United States)

    Smith, Tracey J; Rigassio-Radler, Diane; Denmark, Robert; Haley, Timothy; Touger-Decker, Riva

    2013-06-01

    College students are susceptible to upper respiratory infections (URI) due to inadequate sleep, stress and close living quarters. Certain probiotic strains modulate immune function and may improve health-related quality of life (HRQL) during URI. The present study recruited apparently healthy college students and assessed the effect of probiotics on HRQL outcomes (i.e. self-reported duration, symptom severity and functional impairment of URI) in those who developed URI. Missed school and work days due to URI were also considered. Subjects (n 231) were apparently healthy college students living on campus in residence halls at the Framingham State University (Framingham, MA, USA), and were randomised to receive placebo (n 117) or probiotic-containing powder (daily dose of minimum 1 billion colony-forming units of each Lactobacillus rhamnosus LGG® (LGG®) and Bifidobacterium animalis ssp. lactis BB-12® (BB-12®); n 114) for 12 weeks. Subjects completed The Wisconsin Upper Respiratory Symptom Survey-21 to assess HRQL during URI. The final analyses included 198 subjects (placebo, n 97 and probiotics, n 101). The median duration of URI was significantly shorter by 2 d and median severity score was significantly lower by 34% with probiotics v. placebo (P,0·001), indicating a higher HRQL during URI. Number of missed work days was not different between groups (P=0·429); however, the probiotics group missed significantly fewer school days (mean difference = 0·2 d) compared to the placebo group (P=0·002). LGG® and BB-12® may be beneficial among college students with URI for mitigating decrements in HRQL. More research is warranted regarding mechanisms of action associated with these findings and the cost-benefit of prophylactic supplementation.

  19. Bifidobacterium animalis subsp. lactis BB-12 administration in early childhood: a randomized clinical trial of effects on oral colonization by mutans streptococci and the probiotic.

    Science.gov (United States)

    Taipale, T; Pienihäkkinen, K; Salminen, S; Jokela, J; Söderling, E

    2012-01-01

    A randomized clinical trial studied the effects of early administration of Bifidobacterium animalis subsp. lactis BB-12 (BB-12) on oral colonization of (1) mutans streptococci (MS), and (2) BB-12. In this double-blind, placebo-controlled study, infants (n = 106) received probiotic bacteria (BB-12 group), xylitol (X group), or sorbitol (S group). Test tablets were administered twice a day (from the age of 1-2 months) with a novel slow-release pacifier or a spoon (daily dose of BB-12 10(10) CFU, polyol 200-600 mg). Samples were collected from mucosa/teeth at the age of 8 months and 2 years for BB- 12 determination (qPCR) and plate culturing of MS (MSB, TYCSB), lactobacilli (Rogosa) and yeasts (Sabouraud). The MS levels of the mothers were determined (Dentocult SM Strip Mutans). The baseline characteristics of the three groups were similar. Mean duration of tablet delivery was 14.9 ± 6.7 months. In all groups, >90% of the mothers showed high MS counts (log CFU ≥5). MS colonization percentages of the children at the age of 2 years were rather low (BB-12 group: 6%; X group: 31%; S group: 10%; p < 0.05). The levels of lactobacilli and yeasts did not differ between the groups. BB-12 cell counts barely exceeding the detection limit were found in three of the oral samples of the 8-month-old children; however, the 2-year samples did not contain BB-12. The early administration of BB-12 did not result in permanent oral colonization of this probiotic or significantly affect MS colonization in the children.

  20. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of a health claim related to a combination of Bifidobacterium logum LA 10, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and reducing

    DEFF Research Database (Denmark)

    Tetens, Inge

    substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and reducing intestinal discomfort. The food that is the subject of the health claim is a combination of B. longum LA 101, L...

  1. EFSA NDA Panel (EFSA Panel on Dietetic Products, Nutrition and Allergies), 2014. Scientific Opinion on the substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104

    DEFF Research Database (Denmark)

    Tetens, Inge

    to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and improvement of bowel function by increasing stool frequency. The food that is the subject of the health claim is a combination of four bacterial strains—B. longum...

  2. EFSA NDA Panel (EFSA Panel on Dietetic Products, Nutrition and Allergies), 2014. Scientific Opinion on the substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104

    DEFF Research Database (Denmark)

    Tetens, Inge

    to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and reducing intestinal discomfort. The food that is the subject of the health claim is a combination of four bacterial strains—B. longum LA 101, L. helveticus LA 102...

  3. Spray drying technics of Bifidobacterium lactis%乳酸双歧杆菌喷雾干燥工艺研究

    Institute of Scientific and Technical Information of China (English)

    付博; 马齐; 王卫卫

    2011-01-01

    Spray drying of microencapsulation was used to effectively protect the activity of Bifidobacterium lactis during the production and storage, and technics parameters were also tested. Through wall material selection and orthogonal analysis of relevant parameters, the results showed that the best mass concentration of wall-material was 100 g/L, including β-cyclodextrin and gum acacia, and the mass ratio was 4:6. The mass concentration of core-material was 60 g/L, include trehalose 20 g/L, mannitol 20 g/L, bacteria mire 25 g/L and ascorbic acid 0.01 g/L. The best inlet air temperature was 108±2℃, outlet air temperature was 61±3℃, and tower wall temperature was 50±3℃. Atomizer speed was 17 000 r/min. Under the optimized conditions, the survival bacteria number were over 3.4×109cfu/g, survival rate was 72.12% and encapsulation efficiency was 75.88%.%为有效保证乳酸双歧杆菌在生产、消费及贮藏过程中的菌体活性,利用喷雾干燥法制备双歧杆菌微胶囊,深入研究了喷雾干燥过程中的工艺参数.通过壁材选择及相关工艺参数的正交分析,结果表明:喷雾干燥过程中的最佳壁材质量浓度为100 g/L,包括β-环糊精与阿拉伯树胶粉,且质量比为4∶6,芯材质量浓度60 g/L,包括海藻糖20 g/L,甘露醇20 g/L,菌泥25 g/L和抗坏血酸0.01 g/L.最佳工艺参数为进风温度(108±2)℃,排风温度(61+3)℃,塔壁温度(50+3)℃及雾化器转速17 000 r/min.在此工艺条件下菌粉的活菌数为3.4×109g-1,存活率72.12%.包埋率75.88%.

  4. Improved viability of bifidobacteria in fermented milk by cocultivation with Lactococcus lactis subspecies lactis.

    Science.gov (United States)

    Odamaki, T; Xiao, J Z; Yonezawa, S; Yaeshima, T; Iwatsuki, K

    2011-03-01

    The poor survival of probiotic bacteria in commercial yogurts may limit their potential to exert health benefits in humans. The objective was to improve the survival of bifidobacteria in fermented milk. Cocultivation with some strains of Lactococcus lactis ssp. lactis improved the survival of bifidobacteria in fermented milk during refrigerated storage. Studies on one strain, Lc. lactis ssp. lactis MCC866, showed that the concentrations of dissolved oxygen were kept lower in the cocultivated fermented milk during storage compared with monocultured Bifidobacterium longum BB536 or samples cocultured with another noneffective Lc. lactis ssp. lactis strain. Degradation of genomic DNA was suppressed in the cocultivating system with Lc. lactis ssp. lactis MCC866. Several genes that participated in protection from active oxygen species (e.g., genes coding for alkyl hydroperoxide reductase and Fe(2+) transport system) were expressed at higher levels during refrigerated storage in Lc. lactis ssp. lactis MCC 866 compared with another noneffective Lc. lactis ssp. lactis strain. Concentration of free iron ion was also lower in supernatants of fermented milk cocultivated with B. longum BB536 and Lc. lactis ssp. lactis MCC866. These results suggest that Lc. lactis ssp. lactis MCC 866 is potentially superior in reducing oxygen damage and consequently improves the survival of bifidobacteria in the cocultivating system. This cocultivation system is of industrial interest for producing fermented milk containing viable bifidobacteria with long shelf life.

  5. Scientific Opinion on the substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and improves stool frequency pursuant to Article 13(5 of Regulation (EC No 1924/2006

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA

    2013-02-01

    Full Text Available Following an application from PiLeJe submitted for authorisation of a health claim pursuant to Article 13(5 of Regulation (EC No 1924/2006 via the Competent Authority of France, the Panel on Dietetic Products, Nutrition and Allergies (NDA was asked to deliver an opinion on the scientific substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and improves stools frequency. The food that is the subject of the health claim is a combination of B. longum LA 101, L. helveticus LA 102, L. lactis LA 103 and S. thermophilus LA 104. The information provided was insufficient to establish that the strain L. lactis LA 103 was sufficiently characterised. The Panel considers that if in a combination of several microorganisms and/or ingredients one microorganism or ingredient used in the combination is not sufficiently characterised, then the combination is considered to be not sufficiently characterised. A combination of B. longum LA 101, L. helveticus LA 102, L. lactis LA 103, and S. thermophilus LA 104 is not sufficiently characterised. The Panel concludes that a cause and effect relationship cannot be established between the consumption of a combination of B. longum LA 101, L. helveticus LA 102, L. lactis LA 103 and S. thermophilus LA 104 and improves stool frequency.

  6. Xylo-oligosaccharides alone or in synbiotic combination with Bifidobacterium animalis subsp. lactis induce bifidogenesis and modulate markers of immune function in healthy adults: a double-blind, placebo-controlled, randomised, factorial cross-over study.

    Science.gov (United States)

    Childs, Caroline E; Röytiö, Henna; Alhoniemi, Esa; Fekete, Agnes A; Forssten, Sofia D; Hudjec, Natasa; Lim, Ying Ni; Steger, Cara J; Yaqoob, Parveen; Tuohy, Kieran M; Rastall, Robert A; Ouwehand, Arthur C; Gibson, Glenn R

    2014-06-14

    Prebiotics, probiotics and synbiotics are dietary ingredients with the potential to influence health and mucosal and systemic immune function by altering the composition of the gut microbiota. In the present study, a candidate prebiotic (xylo-oligosaccharide, XOS, 8 g/d), probiotic (Bifidobacterium animalis subsp. lactis Bi-07, 109 colony-forming units (CFU)/d) or synbiotic (8 g XOS+109 CFU Bi-07/d) was given to healthy adults (25-65 years) for 21 d. The aim was to identify the effect of the supplements on bowel habits, self-reported mood, composition of the gut microbiota, blood lipid concentrations and immune function. XOS supplementation increased mean bowel movements per d (P= 0·009), but did not alter the symptoms of bloating, abdominal pain or flatulence or the incidence of any reported adverse events compared with maltodextrin supplementation. XOS supplementation significantly increased participant-reported vitality (P= 0·003) and happiness (P= 0·034). Lowest reported use of analgesics was observed during the XOS+Bi-07 supplementation period (P= 0·004). XOS supplementation significantly increased faecal bifidobacterial counts (P= 0·008) and fasting plasma HDL concentrations (P= 0·005). Bi-07 supplementation significantly increased faecal B. lactis content (P= 0·007), lowered lipopolysaccharide-stimulated IL-4 secretion in whole-blood cultures (P= 0·035) and salivary IgA content (P= 0·040) and increased IL-6 secretion (P= 0·009). XOS supplementation resulted in lower expression of CD16/56 on natural killer T cells (P= 0·027) and lower IL-10 secretion (P= 0·049), while XOS and Bi-07 supplementation reduced the expression of CD19 on B cells (XOS × Bi-07, P= 0·009). The present study demonstrates that XOS induce bifidogenesis, improve aspects of the plasma lipid profile and modulate the markers of immune function in healthy adults. The provision of XOS+Bi-07 as a synbiotic may confer further benefits due to the discrete effects of Bi-07 on the gut

  7. Fermented milk containing Bifidobacterium lactis DN-173 010 improves gastrointestinal well-being and digestive symptoms in women reporting minor digestive symptoms: a randomised, double-blind, parallel, controlled study.

    Science.gov (United States)

    Guyonnet, Denis; Schlumberger, Armelle; Mhamdi, Leila; Jakob, Stefan; Chassany, Olivier

    2009-12-01

    The ability of probiotics to improve bowel habits or transit time has been shown in healthy populations. Additional data are required to support the use of specific probiotics to improve gastrointestinal (GI) well-being. The present study was designed to investigate the effect of consuming fermented milk (FM) on GI well-being, digestive symptoms and health-related quality of life (HRQoL) amongst women without diagnosed GI disorders. In this double-blind, controlled, parallel-design study, subjects were randomised to ingest daily either 2 x 125 g FM containing Bifidobacterium lactis DN-173 010 and yoghurt strains or a control non-fermented dairy product for 4 weeks followed by a 4-week wash-out period. GI well-being and digestive symptoms were assessed weekly. HRQoL was measured every 4 weeks. Data were analysed using analysis of covariance and logistic regression, correcting for baseline values on the full analysis set population of 197 women (aged 18-60 years). The percentage of women reporting an improvement in their GI well-being was significantly (P < 0.01) higher in the FM group v. the control group (OR 1.69; 95 % CI 1.17, 2.45). A significantly (P < 0.05) more pronounced decrease in the composite score of digestive symptoms was observed in the FM group when comparing with the control group (least squares mean - 0.57; 95 % CI - 1.12, - 0.02). Among HRQoL dimensions, the digestive comfort score was significantly (P < 0.05) improved in the FM group compared with the control group. The present study showed that the daily consumption of a specific FM is able to improve GI well-being and digestive symptoms in adult women without GI disorders.

  8. Effect of the probiotic strain Bifidobacterium animalis subsp. lactis, BB-12®, on defecation frequency in healthy subjects with low defecation frequency and abdominal discomfort: a randomised, double-blind, placebo-controlled, parallel-group trial.

    Science.gov (United States)

    Eskesen, Dorte; Jespersen, Lillian; Michelsen, Birgit; Whorwell, Peter J; Müller-Lissner, Stefan; Morberg, Cathrine M

    2015-11-28

    The aim of the present study was to investigate the effect of Bifidobacterium animalis subsp. lactis, BB-12®, on two primary end points - defecation frequency and gastrointestinal (GI) well-being - in healthy adults with low defecation frequency and abdominal discomfort. A total of 1248 subjects were included in a randomised, double-blind, placebo-controlled trial. After a 2-week run-in period, subjects were randomised to 1 or 10 billion colony-forming units/d of the probiotic strain BB-12® or a matching placebo capsule once daily for 4 weeks. Subjects completed a diary on bowel habits, relief of abdominal discomfort and symptoms. GI well-being, defined as global relief of abdominal discomfort, did not show significant differences. The OR for having a defecation frequency above baseline for ≥50% of the time was 1·31 (95% CI 0·98, 1·75), P=0·071, for probiotic treatment overall. Tightening the criteria for being a responder to an increase of ≥1 d/week for ≥50 % of the time resulted in an OR of 1·55 (95% CI 1·22, 1·96), P=0·0003, for treatment overall. A treatment effect on average defecation frequency was found (P=0·0065), with the frequency being significantly higher compared with placebo at all weeks for probiotic treatment overall (all PEffects on defecation frequency were similar for the two doses tested, suggesting that a ceiling effect was reached with the one billion dose. Overall, 4 weeks' supplementation with the probiotic strain BB-12® resulted in a clinically relevant benefit on defecation frequency. The results suggest that consumption of BB-12® improves the GI health of individuals whose symptoms are not sufficiently severe to consult a doctor (ISRCTN18128385).

  9. HOWARU益生菌系列的生理功能研究

    Institute of Scientific and Technical Information of China (English)

    崔岸

    2004-01-01

    本文对二十世纪九十年代新研发的两个市场化的专利益生菌株-HOWARUTM双歧杆菌(Bifidobacterium lactis HN019)和HOARUTM鼠李糖乳杆菌(Lactobacillus rhamnosus HN001)的若干研究进展进行了简要介绍。

  10. Scientific Opinion on the substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and reducing intestinal discomfort pursuant to Article 13(5 of Regulation (EC No 1924/2006

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA

    2014-05-01

    Full Text Available Following an application from PiLeJe, submitted pursuant to Article 13(5 of Regulation (EC No 1924/2006 via the Competent Authority of France, the Panel on Dietetic Products, Nutrition and Allergies (NDA was asked to deliver an opinion on the scientific substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and reducing intestinal discomfort. The food that is the subject of the health claim is a combination of four bacterial strains—B. longum LA 101, L. helveticus LA 102, L. lactis LA 103 and S. thermophilus LA 104. The Panel considers that the food, a combination of B. longum LA 101, L. helveticus LA 102, L. lactis LA 103 and S. thermophilus LA 104, which is the subject of the health claim is sufficiently characterised. The claimed effect proposed by the applicant is "improves intestinal comfort". The Panel considers that reduction of gastro-intestinal discomfort is a beneficial physiological effect. The Panel considers that the only human study provided for the substantiation of the claim (with limitations did not find an effect of a combination of the bacterial strains being the subject of the claim on gastrointestinal discomfort. The Panel concludes that a cause and effect relationship has not been established between the consumption of a combination of B. longum LA 101, L. helveticus LA 102, L. lactis LA 103 and S. thermophilus LA 104 and reducing gastro-intestinal discomfort.

  11. Antimicrobial activity of human β-defensins against lactic acid bacteria.

    Science.gov (United States)

    Wang, Xiao-Fang; Tian, Fei; Cao, Rui-Ming; Li, Jing; Wu, Sheng-Mei; Guo, Xiao-Kui; Chen, Tong-Xin

    2015-01-01

    In this study, we evaluated the antimicrobial activity of human β-defensin-1 (hBD-1), human β-defensin-2 (hBD-2) and human β-defensin-3 (hBD-3) against three internationally common probiotic strains of lactic acid bacterium. Our results indicated that hBD-1, hBD-2 and hBD-3 at the range of 0.08-10 μg/mL do not have obvious antimicrobial activity against these strains. Viability of Bifidobacterium longum JDM301 (B. longum JDM301), Bifidobacterium lactis HN019 (B. lactis HN019) and Lactobacillus rhamnosus GG (LGG) were still very high even at concentration of 10 μg hBD/mL. Then, we explored the mechanism of resistance by using carbonyl cyanide 3-chlorophenylhydrazone (CCCP) to inhibit efflux pumps. In the presence of CCCP, hBD-1, hBD-2 and hBD-3 exhibited enhanced antibacterial effect against B. longum JDM301 and B. lactis HN019, but not against LGG. Efflux pumps in B. longum JDM301 and B. lactis HN019 may partly contribute to their resistance to hBD-1, hBD-2, and hBD-3.

  12. Multiparametric flow cytometry and cell sorting for the assessment of viable, injured, and dead bifidobacterium cells during bile salt stress

    NARCIS (Netherlands)

    Ben Amor, K.; Breeuwer, P.; Verbaarschot, P.; Rombouts, F.M.; Akkermans, A.D.L.; Vos, de W.M.; Abee, T.

    2002-01-01

    Using a flow cytometry-based approach, we assessed the viability of Bifidobacterium lactis DSM 10140 and Bifidobacterium adolescentis DSM 20083 during exposure to bile salt stress. Carboxyfluorescein diacetate (cFDA), propidium iodide (PI), and oxonol [DiBAC4(3)] were used to monitor esterase activi

  13. HOWARU益生菌系列的生理功能研究%Studies on the Physiological Function of HOWARU Probiotics series

    Institute of Scientific and Technical Information of China (English)

    崔岸

    2004-01-01

    本文对二十世纪九十年代新研发的两个市场化的专利益生菌株-HOWARU(tm)双歧杆菌(Bifidobacterium lactis HN019)和HOWARU(tm)鼠李糖乳杆菌(Lactobacillus rhamnosus HN001)的若干研究进展进行了简要介绍.

  14. The GH5 1,4-β-mannanase from Bifidobacterium animalis subsp. lactis Bl-04 possesses a low-affinity mannan-binding module and highlights the diversity of mannanolytic enzymes

    DEFF Research Database (Denmark)

    Morrill, Johan; Kulcinskaja, Evelina; Sulewska, Anna Maria;

    2015-01-01

    β-Mannans are abundant and diverse plant structural and storage polysaccharides. Certain human gut microbiota members including health-promoting Bifidobacterium spp. catabolize dietary mannans. Little insight is available on the enzymology of mannan deconstruction in the gut ecological niche. Here....... Surface plasmon resonance analysis confirmed the binding of the CBM10 to manno-oligosaccharides, albeit with slightly lower affinity than the catalytic module of the enzyme. This is the first example of a low-affinity mannan-specific CBM, which forms a subfamily of CBM10 together with close homologs...... by an exceptionally low Km and the presence of an atypical low affinity CBM, which increases binding to specifically to soluble mannan while causing minimal decrease in catalytic efficiency as opposed to enzymes with canonical mannan binding modules. These features highlight fine tuning of catalytic and binding...

  15. Multiparametric Flow Cytometry and Cell Sorting for the Assessment of Viable, Injured, and Dead Bifidobacterium Cells during Bile Salt Stress

    OpenAIRE

    2002-01-01

    Using a flow cytometry-based approach, we assessed the viability of Bifidobacterium lactis DSM 10140 and Bifidobacterium adolescentis DSM 20083 during exposure to bile salt stress. Carboxyfluorescein diacetate (cFDA), propidium iodide (PI), and oxonol [DiBAC4(3)] were used to monitor esterase activity, membrane integrity, and membrane potential, respectively, as indicators of bacterial viability. Single staining with these probes rapidly and noticeably reflected the behavior of the two strain...

  16. Effects of Lactococcus lactis on composition of intestinal microbiota: Role of nisin

    DEFF Research Database (Denmark)

    Bernbom, Nete; Licht, Tine Rask; Brogren, Carl-Henrik

    2006-01-01

    -producing and the non-nisin-producing L. lactis strains significantly increased the number of Bifidobacterium cells in fecal samples during the first 8 days but decreased the number of enterococci/streptococci in duodenum, ileum, cecum, and colon samples as detected by selective cultivation. No significant changes...

  17. Mupirocin-mucin agar for selective enumeration of Bifidobacterium bifidum.

    Science.gov (United States)

    Pechar, Radko; Rada, Vojtech; Parafati, Lucia; Musilova, Sarka; Bunesova, Vera; Vlkova, Eva; Killer, Jiri; Mrazek, Jakub; Kmet, Vladimir; Svejstil, Roman

    2014-11-17

    Bifidobacterium bifidum is a bacterial species exclusively found in the human intestinal tract. This species is becoming increasingly popular as a probiotic organism added to lyophilized products. In this study, porcine mucin was used as the sole carbon source for the selective enumeration of B. bifidum in probiotic food additives. Thirty-six bifidobacterial strains were cultivated in broth with mucin. Only 13 strains of B. bifidum utilized the mucin to produce acids. B. bifidum was selectively enumerated in eight probiotic food supplements using agar (MM agar) containing mupirocin (100 mg/L) and mucin (20 g/L) as the sole carbon source. MM agar was fully selective if the B. bifidum species was presented together with Bifidobacterium animalis subsp. lactis, Bifidobacterium breve, and Bifidobacterium longum subsp. longum species and with lactic acid bacteria (lactobacilli, streptococci). Isolated strains of B. bifidum were identified using biochemical, PCR, MALDI-TOF procedures and 16S rRNA gene sequencing. The novel selective medium was also suitable for the isolation of B. bifidum strains from human fecal samples.

  18. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of a health claim related to Bifidobacterium animalis subsp. lactis LMG P-21384 and changes in bowel function (ID 2940, further assessment) pursuant to Article 13(1) of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    Following a request from the European Commission, the Panel on Dietetic Products, Nutrition and Allergies (NDA) was asked to provide a scientific opinion on a health claim pursuant to Article 13 of Regulation (EC) No 1924/2006 in the framework of further assessment in relation to Bifidobacterium...

  19. An ATP Binding Cassette Transporter Mediates the Uptake of α-(1,6)-Linked Dietary Oligosaccharides in Bifidobacterium and Correlates with Competitive Growth on These Substrates

    DEFF Research Database (Denmark)

    Hansen, Morten Ejby; Fredslund, Folmer; Andersen, Joakim Mark

    2016-01-01

    and lactobacilli in the human gut. Here we show that the solute binding protein (BlG16BP) associated with an ATP binding cassette (ABC) transporter from the probiotic Bifidobacterium animalis subsp. lactis Bl-04 binds -(1,6)-linked glucosides and galactosides of varying size, linkage, and monosaccharide...

  20. Lysozyme expression in Lactococcus lactis

    NARCIS (Netherlands)

    Guchte, Maarten van de; Wal, Fimme Jan van der; Kok, Jan; Venema, Gerhardus

    1992-01-01

    Three lysozyme-encoding genes, one of eukaryotic and two of prokaryotic origin, were expressed in Lactococcus lactis subsp. lactis. Hen egg white lysozyme (HEL) could be detected in L. lactis lysates by Western blotting. No lysozyme activity was observed, however, presumably because of the absence o

  1. Friendly Bacteria May Lend a Hand to Immune System

    Institute of Scientific and Technical Information of China (English)

    叶朝阳

    2001-01-01

    @@ [选注者言:细菌是可怕的,但是本文标题中的Friendly Bacteria让读者眼睛一亮!读罢全文,我们方才领悟:乳制品中的细菌Bifidobacterium lactis HN019能够在健康成年人体内增强两种免疫系统的细胞.本文核心句是:Certain strains ( 菌系 ) of lactic ( 乳的 ) acid ( 酸 ) bacteria in fermented dairy products may aid digestion, reduce cholesterol ( 胆固醇 ), prevent intestinal infections and boost the immune system, the authors note.

  2. Desenvolvimento de sorvete simbiótico de graviola (Annona muricata L.) com teor reduzido de gordura e avaliação da resistência gastrointestinal dos probióticos in vitro

    OpenAIRE

    Graziela Leal Sousa

    2013-01-01

    O presente trabalho objetivou avaliar o efeito da adição de inulina (I) e a substituição parcial da gordura do leite (G) pelo concentrado de proteína de soro de leite (WPC) sobre a sobrevivência dos probióticos Lactobacillus acidophilus NCFM e Bifidobacterium animalis subsp. lactis HN019 em sorvete de graviola com teor reduzido de gordura, ao longo do período de armazenamento e frente às condições encontradas no trato gastrointestinal (TGI) simuladas in vitro. Adicionalmente, avaliou-se a inf...

  3. Genetic analysis and morphological identification of pilus-like structures in members of the genus Bifidobacterium

    LENUS (Irish Health Repository)

    2011-08-30

    Abstract Background Cell surface pili in Gram positive bacteria have been reported to orchestrate the colonization of host tissues, evasion of immunity and the development of biofilms. So far, little if any information is available on the presence of pilus-like structures in human gut commensals like bifidobacteria. Results and discussion In this report, Atomic Force Microscopy (AFM) of various bifidobacterial strains belonging to Bifidobacterium bifidum, Bifidobacterium longum subsp. longum, Bifidobacterium dentium, Bifidobacterium adolescentis and Bifidobacterium animalis subsp. lactis revealed the existence of appendages resembling pilus-like structures. Interestingly, these microorganisms harbour two to six predicted pilus gene clusters in their genome, with each organized in an operon encompassing the major pilin subunit-encoding gene (designated fimA or fimP) together with one or two minor pilin subunit-encoding genes (designated as fimB and\\/or fimQ), and a gene encoding a sortase enzyme (strA). Quantitative Real Time (qRT)-PCR analysis and RT-PCR experiments revealed a polycistronic mRNA, encompassing the fimA\\/P and fimB\\/Q genes, which are differentially expressed upon cultivation of bifidobacteria on various glycans.

  4. Immune Modulating Capability of Two Exopolysaccharide-Producing Bifidobacterium Strains in a Wistar Rat Model

    Directory of Open Access Journals (Sweden)

    Nuria Salazar

    2014-01-01

    Full Text Available Fermented dairy products are the usual carriers for the delivery of probiotics to humans, Bifidobacterium and Lactobacillus being the most frequently used bacteria. In this work, the strains Bifidobacterium animalis subsp. lactis IPLA R1 and Bifidobacterium longum IPLA E44 were tested for their capability to modulate immune response and the insulin-dependent glucose homeostasis using male Wistar rats fed with a standard diet. Three intervention groups were fed daily for 24 days with 10% skimmed milk, or with 109 cfu of the corresponding strain suspended in the same vehicle. A significant increase of the suppressor-regulatory TGF-β cytokine occurred with both strains in comparison with a control (no intervention group of rats; the highest levels were reached in rats fed IPLA R1. This strain presented an immune protective profile, as it was able to reduce the production of the proinflammatory IL-6. Moreover, phosphorylated Akt kinase decreased in gastroctemius muscle of rats fed the strain IPLA R1, without affecting the glucose, insulin, and HOMA index in blood, or levels of Glut-4 located in the membrane of muscle and adipose tissue cells. Therefore, the strain B. animalis subsp. lactis IPLA R1 is a probiotic candidate to be tested in mild grade inflammation animal models.

  5. Use of modified Lactobacillus selective medium and Bifidobacterium iodoacetate medium for differential enumeration of Lactobacillus acidophilus and Bifidobacterium spp. in powdered nutritional products.

    Science.gov (United States)

    Ingham, S C

    1999-01-01

    Modified Lactobacillus selective agar (APT agar + sodium acetate and glacial acetic acid; mLBS) was compared to selective modified Lactobacillus selective medium (LBS agar + tomato juice and acetic acid; mLSM) and nonselective de Man Rogosa Sharpe (MRS) agar for the enumeration of Lactobacillus acidophilus in probiotic-containing powdered nutritional products. The mLBS agar was equivalent to MRS agar and superior to the mLSM agar for enumerating L. acidophilus in products stored in sealed cans at 22 degrees C. When samples were analyzed for L. acidophilus concentration after high temperature storage in sealed cans or storage in open cans at high relative humidity, the mLBS and MRS agars were highly correlated (r2 = 0.93). Modified Bifidobacterium iodoacetate medium (12.5 mg iodoacetic acid/liter; mBIM) was compared to MRS agar + bile, cysteine, and dicloxacillin (MRS + BCD) for enumerating Bifidobacterium infantis or Bifidobacterium lactis in the nutritional products. The two media were equivalent for enumerating B. infantis in product stored at 22 degrees C in sealed cans. However, the two media were poorly correlated (r2nutritional products stored under high temperature and/or high relative humidity conditions.

  6. Construction and Expression of β-galactosidase Genetically Engineered Lactococcus lactis

    Institute of Scientific and Technical Information of China (English)

    吕晓英; 张朝武; 裴晓方; 刘祥; 余倩; 刘衡川

    2004-01-01

    Our objective is to solve the lactose malabsorption and intolerance of human beings by combining mlcro-ecology path with genetic engineering technique. Plasmid pMG36e was used to clone and express a β-galactosidase gene from L.delbrueckii bulgaricus strain 1. 1480 in the Lactococcus lactis subsp, cremoris MG1363 and Lactococcus lactis subsp. lactis IL1403. The recombinant plasmid was preserved and proliferated in Escherichia coli ( E. coli) JM109, and transformed into MG1363 and 1L1403 by electroporation. The protein expression was studied. (1) The bifidobacterium culture medium (BBL) was suitable for the growth of the strain 1. 1480. (2) With 13 amino acids at the N-terminus from the vector, β-galactosidase fusion protein (which retained the enzyme activity) could be successfully expressed in E. coli JM109, MG1363 and IL1403, but the expression quantity was larger in the former than in the latter two. (3) The SD sequence designed could be successfully recognized by both the E. coli and the Lactococcus lactis, but the expression level of the non-fusion β-galac-tosidase protein was lower than that of the fusion protein in the same host. The β-galactosidase genetically engineered E.coli JM109 is a useful tool to produce this enzyme in vitro. The signal peptide of the usp45 protein from the Lactococcus lactis can be added before the promoter sequence to promote β-galactosidase secretion from Lactococcus lactis. The potential application of the β-galactosidase genetically engineered MG1363 and IL1403 to cure the lactose malabsorption and lactose intolerance in both health food and medicine is promising。

  7. Enhancement of Nisin Production by Lactococcus lactis subsp. lactis.

    Science.gov (United States)

    Dussault, Dominic; Vu, Khanh Dang; Lacroix, Monique

    2016-09-01

    Lactococcus lactis subsp lactis BSA (L. lactis BSA) was isolated from a commercial fermented product (BSA Food Ingredients, Montreal, Canada) containing mixed bacteria that are used as starter for food fermentation. In order to increase the bacteriocin production by L. lactis BSA, different fermentation conditions were conducted. They included different volumetric combinations of two culture media (the Man, Rogosa and Sharpe (MRS) broth and skim milk), agitation level (0 and 100 rpm) and concentration of commercial nisin (0, 0.15, and 0.30 µg/ml) added into culture media as stimulant agent for nisin production. During fermentation, samples were collected and used for antibacterial evaluation against Lactobacillus sakei using agar diffusion assay. Results showed that medium containing 50 % MRS broth and 50 % skim milk gave better antibacterial activity as compared to other medium formulations. Agitation (100 rpm) did not improve nisin production by L. lactis BSA. Adding 0.15 µg/ml of nisin into the medium-containing 50 % MRS broth and 50 % skim milk caused the highest nisin activity of 18,820 AU/ml as compared to other medium formulations. This activity was 4 and ~3 times higher than medium containing 100 % MRS broth without added nisin (~4700 AU/ml) and 100 % MRS broth with 0.15 µg/ml of added nisin (~6650 AU/ml), respectively.

  8. Effect of Bifidobacterium upon Clostridium difficile growth and toxicity when co-cultured in different prebiotic substrates

    Directory of Open Access Journals (Sweden)

    Lorena Valdés Varela

    2016-05-01

    Full Text Available The intestinal overgrowth of Clostridium difficile, often after disturbance of the gut microbiota by antibiotic treatment, leads to C. difficile infection (CDI which manifestation ranges from mild diarrhoea to life-threatening conditions. The increasing CDI incidence, not only in compromised subjects but also in traditionally considered low-risk populations, together with the frequent relapses of the disease, has attracted the interest for prevention/therapeutic options. Among these, probiotics, prebiotics or synbiotics constitute a promising approach. In this study we determined the potential of selected Bifidobacterium strains for the inhibition of C. difficile growth and toxicity in different carbon sources. We conducted co-cultures of the toxigenic strain C. difficile LMG21717 with four Bifidobacterium strains (Bifidobacterium longum IPLA20022, Bifidobacterium breve IPLA20006, Bifidobacterium bifidum IPLA20015, and Bifidobacterium animalis subsp. lactis Bb12 in the presence of various prebiotic substrates (Inulin, Synergy and Actilight or glucose, and compared the results with those obtained for the corresponding mono-cultures. C. difficile and bifidobacteria levels were quantified by qPCR; the pH and the production of short chain fatty acids was also determined. Moreover, supernatants of the cultures were collected to evaluate their toxicity using a recently developed model. Results showed that co-culture with B. longum IPLA20022 and B. breve IPLA20006 in the presence of short-chain fructooligosaccharides, but not of Inulin, as carbon source significantly reduced the growth of the pathogen. With the sole exception of B. animalis Bb12, whose growth was enhanced, the presence of C. difficile did not show major effects upon the growth of the bifidobacteria. In accordance with the growth data, B. longum and B. breve were the strains showing higher reduction in the toxicity of the co-culture supernatants.

  9. FORMULASI PRODUK SUSU FERMENTASI KERING DENGAN PENAMBAHAN BAKTERI PROBIOTIK Lactobacillus casei DAN Bifidobacterium longum

    Directory of Open Access Journals (Sweden)

    Dida Hani Rahman

    2012-11-01

    Full Text Available ABSTRACTFermented milk is a healthy product that has many benefits especially for human digestive tract. Manufacturing of probiotic fermented milk products as a functional food with a viable long shelf life needs to be developed. The purpose of this study was to formulate a dried fermented milk product using probiotic bacterias. The experimental design study was a complete random design with 4 treatments using different lactic acid bacteria (LAB: A1 (Streptococcus lactis: 0.5%; A2 (Streptococcus lactis: 0.25% and Lactobacillus casei: 0.25%; A3 (Streptococcus lactis: 0.25%, Lactobacillus bulgaricus: 0.125%, Streptococcus thermophiles: 0.125%; and A4 (Streptococcus lactis: 0.25% and Bifidobacterium longum: 0.25%. The highest level of hardness was A2 product and the highest level of tenderness is A1 product. Results of proximate analysis showed that dried fermented milk products had high levels of the protein, calcium, and phosphorus. Microbiological test results showed that the amount of lactic acid bacteria (BAL in dried fermented milk products were eligible based on CODEX: 243 (2003. Statictical analysis using ANOVA in the hedonic quality test showed that the treatments were significantly different (p 0.05 on the attributes of color, aroma, hardness, and flavor except texture.Key words: dried fermented milk, functional food, probiotic, lactic acid bacteria (LABABSTRAKSusu fermentasi merupakan produk kesehatan yang mempunyai banyak manfaat terutama untuk saluran pencernaan manusia. Pembuatan produk susu fermentasi probiotik sebagai makanan fungsional yang mempunyai kelangsungan hidup dan daya simpan yang lama perlu dikembangan. Tujuan penelitian ini adalah untuk memformulasikan susu fermentasi kering menggunakan bakteri probiotik. Desain penelitian yang digunakan adalah rancangan acak lengkap dengan 4 perlakuan menggunakan 4 bakteri asam laktat (BAL, (Streptococcus lactis, 0.5%, A2 (Streptococcus lactis; 0.25% dan Lactobacillus casei; 0.25%, A3

  10. Performances of new isolates of Bifidobacterium on fermentation of soymilk.

    Science.gov (United States)

    Havas, Petra; Kun, Szilárd; Perger-Mészáros, Izabell; Rezessy-Szabó, Judit M; Nguyen, Quang D

    2015-12-01

    Growth and metabolic activity of several new, human origin isolates of Bifidobacterium strains were investigated. All tested bifidobacteria strains were grown well on the native soymilk medium without any additional nutrients. The fermentation processes cultured with initial cell concentrations in 10⁵ -10⁷ cfu/ml resulted in 10⁸ cfu/ml after 8-12 h of incubation in soymilk, and were kept viable up to the end of fermentation (48 h). Volumetric productivities of B. bifidum B3.2, B. bifidum B7.1 and B. breve B9.14 were 1.6 × 10¹⁰ cfu/L.h, 4.5 × 10¹⁰ cfu/L.h and 7.6 × 10⁹ cfu/L.h, respectively, whereas these values of B. lactis Bb-12 and B. longum Bb-46 probiotic strains were 2.7 × 10⁹ cfu/L.h and 1.0 x 10¹⁰ cfu/L.h. The α-galactosidase activities were also detected in the intracellular fraction of the disrupted cells. Productions of lactic and acetic acids were in the range of 23-60 mmol/L and 2.4-5.6 mmol/L, respectively. Molar ratios of acetate to lactate in all tested strains varied from 0.05-0.1 that are very promising for further technological development of probiotic fermented soy-based food products.

  11. Genome Sequence of Lactococcus lactis subsp. lactis NCDO 2118, a GABA-Producing Strain

    DEFF Research Database (Denmark)

    Oliveira, Letícia C; Saraiva, Tessália D L; Soares, Siomar C

    2014-01-01

    Lactococcus lactis subsp. lactis NCDO 2118 is a nondairy lactic acid bacterium, a xylose fermenter, and a gamma-aminobutyric acid (GABA) producer isolated from frozen peas. Here, we report the complete genome sequence of L. lactis NCDO 2118, a strain with probiotic potential activity....

  12. Immune effects of the probiotic Bifidobacterium breve

    NARCIS (Netherlands)

    Ezendam J; van Loveren H; TOX

    2007-01-01

    Bifidobacterium breve, een probiotische bacterie, heeft gunstige effecten op zowel allergieen als autoimmuniteit - een afweerreactie op lichaamseigen bestanddelen - bij proefdieren. Probiotica worden in reclameboodschappen ook wel 'goede bacterien' genoemd. Fabrikanten claimen een positi

  13. The Lactococcus lactis Thioredoxin System

    DEFF Research Database (Denmark)

    Efler, Petr

    -dependent thioredoxin reductase (NTR) in order to complete its catalytic cycle. Glutathione-dependent glutaredoxin complements Trx in many organisms. This thesis focuses on disulfide reduction pathways in Lactococcus lactis, an important industrial microorganism used traditionally for cheese and buttermilk production...... ribonucleotide reductase (NrdEF). Physiological functions of LlTrxA and LlTrxD were studied using ΔtrxA, ΔtrxD and ΔtrxAΔtrxD mutant strains of L. lactis ssp. cremoris MG1363 exposed to various stress conditions and comparing them to the wild type (wt) strain. These experiments revealed that the ΔtrxA genotype...... with a previous study showing that NTR in L. lactis is not essential. Therefore, the presence of an additional thiol redox system is hypothesized. Biochemical studies demonstrated that recombinant LlTrxA, LlTrxD and LlNrdH are substrates for LlNTR, while only LlTrxA and LlNrdH are efficiently reduced by E. coli...

  14. Microbiota of Minas cheese as influenced by the nisin producer Lactococcus lactis subsp. lactis GLc05.

    Science.gov (United States)

    Perin, Luana Martins; Dal Bello, Barbara; Belviso, Simona; Zeppa, Giuseppe; de Carvalho, Antônio Fernandes; Cocolin, Luca; Nero, Luís Augusto

    2015-12-01

    Minas cheese is a popular dairy product in Brazil that is traditionally produced using raw or pasteurized cow milk. This study proposed an alternative production of Minas cheese using raw goat milk added of a nisin producer Lactococcus lactis subsp. lactis GLc05. An in situ investigation was carried on to evaluate the interactions between the L. lactis subsp. lactis GLc05 and the autochthonous microbiota of a Minas cheese during the ripening; production of biogenic amines (BAs) was assessed as a safety aspect. Minas cheese was produced in two treatments (A, by adding L. lactis subsp. lactis GLc05, and B, without adding this strain), in three independent repetitions (R1, R2, and R3). Culture dependent (direct plating) and independent (rep-PCR and PCR-DGGE) methods were employed to characterize the microbiota and to assess the possible interferences caused by L. lactis subsp. lactis GLc05. BA amounts were measured using HPLC. A significant decrease in coagulase-positive cocci was observed in the cheeses produced by adding L. lactis subsp. lactis GLc05 (cheese A). The rep-PCR and PCR-DGGE highlighted the differences in the microbiota of both cheeses, separating them into two different clusters. Lactococcus sp. was found as the main microorganism in both cheeses, and the microbiota of cheese A presented a higher number of species. High concentrations of tyramine were found in both cheeses and, at specific ripening times, the BA amounts in cheese B were significantly higher than in cheese A (pnisin producer L. lactis subsp. lactis GLc05 was demonstrated in situ, by demonstration of its influence in the complex microbiota naturally present in a raw goat milk cheese and by controlling the growth of coagulase-positive cocci. L. lactis subsp. lactis GLc05 influenced also the production of BA determining that their amounts in the cheeses were maintained at acceptable levels for human consumption.

  15. Bifidobacterium reuteri sp. nov., Bifidobacterium callitrichos sp. nov., Bifidobacterium saguini sp. nov., Bifidobacterium stellenboschense sp. nov. and Bifidobacterium biavatii sp. nov. isolated from faeces of common marmoset (Callithrix jacchus) and red-handed tamarin (Saguinus midas).

    Science.gov (United States)

    Endo, Akihito; Futagawa-Endo, Yuka; Schumann, Peter; Pukall, Rüdiger; Dicks, Leon M T

    2012-03-01

    Five strains of bifidobacteria were isolated from faeces of a common marmoset (Callithrix jacchus) and a red-handed tamarin (Saguinus midas). The five isolates clustered inside the phylogenetic group of the genus Bifidobacterium but did not show high sequence similarities between the isolates and to known species in the genus by phylogenetic analysis based on 16S rRNA gene sequences. Sequence analyses of dnaJ1 and hsp60 also indicated their independent phylogenetic positions to each other in the Bifidobacterium cluster. DNA G+C contents of the species ranged from 57.3 to 66.3 mol%, which is within the values recorded for Bifidobacterium species. All isolates showed fructose-6-phosphate phosphoketolase activity. Based on the data provided, the five isolates represent five novel species, for which the names Bifidobacterium reuteri sp. nov. (type strain: AFB22-1(T) = JCM 17295(T) = DSM 23975(T)), Bifidobacterium callitrichos sp. nov. (type strain: AFB22-5(T) = JCM 17296(T) = DSM 23973(T)), Bifidobacterium saguini sp. nov. (type strain: AFB23-1(T) = JCM 17297(T) = DSM 23967(T)), Bifidobacterium stellenboschense sp. nov. (type strain: AFB23-3(T) = JCM 17298(T) = DSM 23968(T)) and Bifidobacterium biavatii sp. nov. (type strain: AFB23-4(T) = JCM 17299(T) = DSM 23969(T)) are proposed.

  16. Multiparametric flow cytometry and cell sorting for the assessment of viable, injured, and dead bifidobacterium cells during bile salt stress.

    Science.gov (United States)

    Amor, Kaouther Ben; Breeuwer, Pieter; Verbaarschot, Patrick; Rombouts, Frank M; Akkermans, Antoon D L; De Vos, Willem M; Abee, Tjakko

    2002-11-01

    Using a flow cytometry-based approach, we assessed the viability of Bifidobacterium lactis DSM 10140 and Bifidobacterium adolescentis DSM 20083 during exposure to bile salt stress. Carboxyfluorescein diacetate (cFDA), propidium iodide (PI), and oxonol [DiBAC4(3)] were used to monitor esterase activity, membrane integrity, and membrane potential, respectively, as indicators of bacterial viability. Single staining with these probes rapidly and noticeably reflected the behavior of the two strains during stress exposure. However, the flow cytometry results tended to overestimate the viability of the two strains compared to plate counts, which appeared to be related to the nonculturability of a fraction of the population as a result of sublethal injury caused by bile salts. When the cells were simultaneously stained with cFDA and PI, flow cytometry and cell sorting revealed a striking physiological heterogeneity within the stressed bifidobacterium population. Three subpopulations could be identified based on their differential uptake of the probes: cF-stained, cF and PI double-stained, and PI-stained subpopulations, representing viable, injured, and dead cells, respectively. Following sorting and recovery, a significant fraction of the double-stained subpopulation (40%) could resume growth on agar plates. Our results show that in situ assessment of the physiological activity of stressed bifidobacteria using multiparameter flow cytometry and cell sorting may provide a powerful and sensitive tool for assessment of the viability and stability of probiotics.

  17. A proliferative probiotic Bifidobacterium strain in the gut ameliorates progression of metabolic disorders via microbiota modulation and acetate elevation

    Science.gov (United States)

    Aoki, Ryo; Kamikado, Kohei; Suda, Wataru; Takii, Hiroshi; Mikami, Yumiko; Suganuma, Natsuki; Hattori, Masahira; Koga, Yasuhiro

    2017-01-01

    The gut microbiota is an important contributor to the worldwide prevalence of metabolic syndrome (MS), which includes obesity and diabetes. The anti-MS effects exerted by Bifidobacterium animalis ssp. lactis GCL2505 (BlaG), a highly proliferative Bifidobacterium strain in the gut, and B. longum ssp. longum JCM1217T (BloJ) were comparatively examined. BlaG treatment reduced visceral fat accumulation and improved glucose tolerance, whereas BloJ had no effect on these parameters. Gut microbial analysis revealed that BlaG exerted stronger effects on the overall bacterial structure of the gut microbiota than BloJ, including enrichment of the genus Bifidobacterium. The levels of acetate and glucagon-like peptide-1 were increased by BlaG treatment in both the gut and plasma, but not by BloJ treatment. Correlation analysis suggested that the elevation of gut acetate levels by BlaG treatment plays a pivotal role in the BlaG-induced anti-MS effects. These findings indicated that BlaG, a highly viable and proliferative probiotic, improves metabolic disorders by modulating gut microbiota, which results in the elevation of SCFAs, especially acetate. PMID:28252037

  18. Enhancing bile tolerance improves survival and persistence of Bifidobacterium and Lactococcus in the murine gastrointestinal tract

    Directory of Open Access Journals (Sweden)

    Hill Colin

    2008-10-01

    Full Text Available Abstract Background The majority of commensal gastrointestinal bacteria used as probiotics are highly adapted to the specialised environment of the large bowel. However, unlike pathogenic bacteria; they are often inadequately equipped to endure the physicochemical stresses of gastrointestinal (GI delivery in the host. Herein we outline a patho-biotechnology strategy to improve gastric delivery and host adaptation of a probiotic strain Bifidobacterium breve UCC2003 and the generally regarded as safe (GRAS organism Lactococcus lactis NZ9000. Results In vitro bile tolerance of both strains was significantly enhanced (P Listeria monocytogenes bile resistance mechanism BilE. Strains harbouring bilE were also recovered at significantly higher levels (P n = 5, following oral inoculation. Furthermore, a B. breve strain expressing bilE demonstrated increased efficacy relative to the wild-type strain in reducing oral L. monocytogenes infection in mice. Conclusion Collectively the data indicates that bile tolerance can be enhanced in Bifidobacterium and Lactococcus species through rational genetic manipulation and that this can significantly improve delivery to and colonisation of the GI tract.

  19. Tulum Peynirlerinden izole Edilen Lactococcus lactis subsp. lactis YBML9 ve

    Directory of Open Access Journals (Sweden)

    Yasin TUNCER

    2009-04-01

    Full Text Available Bu çalısmanın amacı tulum peynirlerinden izole edilen Lactococcus lactis suslarının fenotipik tanısı ve bu suslar tarafından üretilen bakteriyosinlerin kısmi karakterizasyonlarıdır. Bu amaçla Türkiye'nin sekiz farklı ilinden (Ankara, Antalya, Burdur, Denizli, Erzincan, Isparta, İstanbul ve İzmir yöresel pazarlardan toplanan 60 adet tulum peyniri örneginden 40 adet Lactococcus lactis susu (31 adet L. lactis subsp. lactis ve 9 adet L. lactis subsp. cremoris izole edildi. 40 adet L. lactis susu içerisinden, 2 adet L. lactis subsp. lactis (YBML9 ve YBML21 susu bakteriyosin üretme yeteneginde bulundu. L. lactis subsp. lactis YBML9 ve YBML21 susları tarafından üretilen bakteriyosinler, farklı enzim, pH ve sıcaklık uygulamaları sonucu; sırasıyla nisin ve laktisin 481 olarak tanımlandı.

  20. Structural basis for arabinoxylo‐oligosaccharide capture by the probiotic Bifidobacterium animalis subsp. lactis Bl‐04

    DEFF Research Database (Denmark)

    Hansen, Morten Ejby; Fredslund, Folmer; Vujicic‐Zagar, Andreja

    2013-01-01

    specificity for arabinosyl‐decorated and undecorated xylo‐oligosaccharides, with preference for tri‐ and tetra‐saccharides. Crystal structures of BlAXBP in complex with four different ligands revealed the basis for this versatility. Uniquely, the protein was able to recognize oligosaccharides in two opposite...

  1. Elucidating Flux Regulation of the Fermentation Modes of Lactococcus lactis

    DEFF Research Database (Denmark)

    Chan, Siu Hung Joshua

    The long history of application to the dairy industry has established Lactococcus lactis (L. lactis), the lactic acid bacterium, as one of the most extensively characterized low GC organisms. The relatively simple metabolism of L. lactis has also made it an attractive target for metabolic enginee...... method for combining metabolic flux analysis and elementary mode analysis was developed and applied to analyse a case of amino acid metabolism of L. lactis.......The long history of application to the dairy industry has established Lactococcus lactis (L. lactis), the lactic acid bacterium, as one of the most extensively characterized low GC organisms. The relatively simple metabolism of L. lactis has also made it an attractive target for metabolic...... an important subject for basic research in cellular metabolism because L. lactis exhibits an interesting metabolic shift. Under anaerobic conditions, on fast fermentable sugars, L. lactis produces lactate as the primary product, known as homolactic fermentation but on slowly fermentable sugars, significant...

  2. A β1-6/β1-3 galactosidase from B ifidobacterium animalis subsp. lactis Bl-04 gives insight into sub-specificities of β-galactoside catabolism within B ifidobacterium

    DEFF Research Database (Denmark)

    Viborg, Alexander Holm; Fredslund, Folmer; Katayama, Takane

    2014-01-01

    The Bifidobacterium genus harbours several health promoting members of the gut microbiota. Bifidobacteria display metabolic specialization by preferentially utilizing dietary or host-derived β-galactosides. This study investigates the biochemistry and structure of a glycoside hydrolase family 42...... (GH42) β-galactosidase from the probiotic Bifidobacterium animalis subsp. lactis Bl-04 (BlGal42A). BlGal42A displays a preference for undecorated β1-6 and β1-3 linked galactosides and populates a phylogenetic cluster with close bifidobacterial homologues implicated in the utilization of N...

  3. Comparative genomics of the Bifidobacterium breve taxon

    NARCIS (Netherlands)

    Bottacini, Francesca; O'Connell Motherway, Mary; Kuczynski, Justin; O'Connell, Kerry Joan; Serafini, Fausta; Duranti, Sabrina; Milani, Christian; Turroni, Francesca; Lugli, Gabriele Andrea; Zomer, Aldert; Zhurina, Daria; Riedel, Christian; Ventura, Marco; van Sinderen, Douwe

    2014-01-01

    BACKGROUND: Bifidobacteria are commonly found as part of the microbiota of the gastrointestinal tract (GIT) of a broad range of hosts, where their presence is positively correlated with the host's health status. In this study, we assessed the genomes of thirteen representatives of Bifidobacterium br

  4. High efficiency electrotransformation of Lactococcus lactis spp. lactis cells pretreated with lithium acetate and dithiothreitol

    Directory of Open Access Journals (Sweden)

    Filioussis George

    2007-03-01

    Full Text Available Abstract Background A goal for the food industry has always been to improve strains of Lactococcus lactis and stabilize beneficial traits. Genetic engineering is used extensively for manipulating this lactic acid bacterium, while electropolation is the most widely used technique for introducing foreign DNA into cells. The efficiency of electrotransformation depends on the level of electropermealization and pretreatment with chemicals which alter cell wall permeability, resulting in improved transformation efficiencies is rather common practice in bacteria as in yeasts and fungi. In the present study, treatment with lithium acetate (LiAc and dithiothreitol (DTT in various combinations was applied to L. lactis spp. lactis cells of the early-log phase prior to electroporation with plasmid pTRKH3 (a 7.8 kb shuttle vector, suitable for cloning into L. lactis. Two strains of L. lactis spp. lactis were used, L. lactis spp. lactis LM0230 and ATCC 11454. To the best of our knowledge these agents have never been used before with L. lactis or other bacteria. Results Electrotransformation efficiencies of up to 105 transformants per μg DNA have been reported in the literature for L. lactis spp.lactis LM0230. We report here that treatment with LiAc and DDT before electroporation increased transformation efficiency to 225 ± 52.5 × 107 transformants per μg DNA, while with untreated cells or treated with LiAc alone transformation efficiency approximated 1.2 ± 0.5 × 105 transformants per μg DNA. Results of the same trend were obtained with L. lactis ATCC 11454, although transformation efficiency of this strain was significantly lower. No difference was found in the survival rate of pretreated cells after electroporation. Transformation efficiency was found to vary directly with cell density and that of 1010 cells/ml resulted in the highest efficiencies. Following electrotransformation of pretreated cells with LiAc and DDT, pTRKH3 stability was examined

  5. Lactococcus lactis expressing food-grade β-galactosidase alleviates lactose intolerance symptoms in post-weaning Balb/c mice.

    Science.gov (United States)

    Li, Jingjie; Zhang, Wen; Wang, Chuan; Yu, Qian; Dai, Ruirui; Pei, Xiaofang

    2012-12-01

    The endogenous β-galactosidase expressed in intestinal microbes is demonstrated to help humans in lactose usage, and treatment associated with the promotion of beneficial microorganism in the gut is correlated with lactose tolerance. From this point, a kind of recombinant live β-galactosidase delivery system using food-grade protein expression techniques and selected probiotics as vehicle was promoted by us for the purpose of application in lactose intolerance subjects. Previously, a recombinant Lactococcus lactis MG1363 strain expressing food-grade β-galactosidase, the L. lactis MG1363/FGZW, was successfully constructed and evaluated in vitro. This study was conducted to in vivo evaluate its efficacy on alleviating lactose intolerance symptoms in post-weaning Balb/c mice, which were orally administered with 1 × 10⁶ CFU or 1 × 10⁸ CFU of L. lactis MG1363/FGZW daily for 4 weeks before lactose challenge. In comparison with naïve mice, the mice administered with L. lactis MG1363/FGZW showed significant alleviation of diarrhea symptoms in less total feces weight within 6 h post-challenge and suppressed intestinal motility after lactose challenge, although there was no significant increase of β-galactosidase activity in small intestine. The alleviation also correlated with higher species abundance, more Bifidobacterium colonization, and stronger colonization resistance in mice intestinal microflora. Therefore, this recombinant L. lactis strain effectively alleviated diarrhea symptom induced by lactose uptake in lactose intolerance model mice with the probable mechanism of promotion of lactic acid bacteria to differentiate and predominantly colonize in gut microbial community, thus making it a promising probiotic for lactose intolerance subjects.

  6. Draft Genome Sequence of Lactococcus lactis subsp. lactis bv. diacetylactis CRL264, a Citrate-Fermenting Strain

    Science.gov (United States)

    Zuljan, Federico; Espariz, Martín; Blancato, Victor S.; Esteban, Luis; Alarcón, Sergio

    2016-01-01

    We report the draft genome sequence of Lactococcus lactis subsp. lactis bv. diacetylactis CRL264, a natural strain isolated from artisanal cheese from northwest Argentina. L. lactis subsp. lactis bv. diacetylactis is one of the most important microorganisms used as starter culture around the world. The CRL264 strain constitutes a model microorganism in the studies on the generation of aroma compounds (diacetyl, acetoin, and 2,3-butanediol) by lactic acid bacteria. Our genome analysis shows similar genetic organization to other available genomes of L. lactis bv. diacetylactis strains. PMID:26847906

  7. Purification and partial characterization of bacteriocin produced by Lactococcus lactis ssp. lactis LL171.

    Science.gov (United States)

    Kumari, Archana; Akkoç, Nefise; Akçelik, Mustafa

    2012-04-01

    Lactic acid bacteria (LAB) are possessing ability to synthesize antimicrobial compounds (like bacteriocin) during their growth. In this regard, novel bacteriocin compound secreting capability of LAB isolated from Tulum Cheese in Turkey was demonstrated. The synthesized bacteriocin was purified by ammonium sulphate precipitation, dialysis and gel filtration. The molecular weight (≈3.4 kDa) of obtained bacteriocin was confirmed by SDS-PAGE, which revealed single peptide band. Molecular identification of LAB strain isolated from Tulum Cheese was conducted using 16S rDNA gene sequencing as Lactococcus lactis ssp. lactis LL171. The amino acid sequences (KKIDTRTGKTMEKTEKKIELSLKNMKTAT) of the bacteriocin from Lactococcus lactis ssp. lactis LL171 was found unique and novel than reported bacteriocins. Further, the bacteriocin was possessed the thermostable property and active at wide range of pH values from 1 to 11. Thus, bacteriocin reported in this study has the potential applications property as food preservative agent.

  8. Bacteriocinogenic Lactococcus lactis subsp. lactis DF04Mi isolated from goat milk: characterization of the bacteriocin.

    Science.gov (United States)

    Furtado, Danielle N; Todorov, Svetoslav D; Landgraf, Mariza; Destro, Maria T; Franco, Bernadette D G M

    2014-01-01

    Lactic acid bacteria capable of producing bacteriocins and presenting probiotic potential open innovative technological applications in the dairy industry. In this study, a bacteriocinogenic strain (Lactococcus lactis subsp. lactis DF4Mi) was isolated from goat milk, and studied for its antimicrobial activity. The bacteriocin presented a broad spectrum of activity, was sensitive to proteolytic enzymes, resistant to heat and pH extremes, and not affected by the presence of SDS, Tween 20, Tween 80, EDTA or NaCl. Bacteriocin production was dependent on the components of the culture media, especially nitrogen source and salts. When tested by PCR, the bacteriocin gene presented 100% homology to nisin Z gene. These properties indicate that this L. lactis subsp. lactis DF4Mi can be used for enhancement of dairy foods safety and quality.

  9. Bacteriocinogenic Lactococcus lactis subsp. lactis DF04Mi isolated from goat milk: characterization of the bacteriocin

    Directory of Open Access Journals (Sweden)

    Danielle N. Furtado

    2014-12-01

    Full Text Available Lactic acid bacteria capable of producing bacteriocins and presenting probiotic potential open innovative technological applications in the dairy industry. In this study, a bacteriocinogenic strain (Lactococcus lactis subsp. lactis DF4Mi was isolated from goat milk, and studied for its antimicrobial activity. The bacteriocin presented a broad spectrum of activity, was sensitive to proteolytic enzymes, resistant to heat and pH extremes, and not affected by the presence of SDS, Tween 20, Tween 80, EDTA or NaCl. Bacteriocin production was dependent on the components of the culture media, especially nitrogen source and salts. When tested by PCR, the bacteriocin gene presented 100% homology to nisin Z gene. These properties indicate that this L. lactis subsp. lactis DF4Mi can be used for enhancement of dairy foods safety and quality.

  10. Control of Brochothrix thermosphacta in pork meat using Lactococcus lactis subsp. lactis I23 isolated from beef

    Directory of Open Access Journals (Sweden)

    Olusegun A Olaoye

    2015-06-01

    Full Text Available This study evaluated the antimicrobial activities of two lactic acid bacteria (LAB Lactococcus lactis subsp. lactis I23 and L. lactis subsp. hordinae E91 against Brochothrix thermosphacta in pork during storage at ambient temperature (30oC over 7 days. Both the LAB strains and spoilage organism were inoculated on fresh pork samples at 1x106cfu/g. About 3 log reduction in the spoilage organism was obtained in LAB treated samples after 48 h of storage. The spoilage organism was confirmed to be sensitive to the bacteriocin nisin produced by Lactococcus lactis subsp. lactis I23. There were reductions in the counts of Salmonella typhimurium, Listeria monocytogenes, Enterobacteriaceae and Staphylococcus in the treated samples. Conclusively, growth of B. thermosphacta could be effectively controlled by nisin producing Lactococcus lactis subsp. lactis I23 in fresh pork during storage, thereby enhancing shelf life of the product.

  11. Bile salt hydrolase of Bifidobacterium longum - Biochemical and genetic characterization

    NARCIS (Netherlands)

    Tanaka, H; Hashiba, Honoo; Kok, Jan; Mierau, Igor

    2000-01-01

    A bile salt hydrolase (BSH) was isolated from Bifidobacterium longum SBT2928, purified, and characterized, Furthermore, we describe for the first time cloning and analysis of the gene encoding BSII (bsh) in a member of the genus Bifidobacterium. The enzyme has a native molecular weight of 125,000 to

  12. Protein export elements from Lactococcus lactis

    NARCIS (Netherlands)

    Perez-Martinez, Gaspar; Kok, Jan; Venema, Gerhardus; Dijl, Jan Maarten van; Smith, Hilda; Bron, Sierd

    1992-01-01

    Broad-host-range plasmids carrying α-amylase or β-lactamase reporter genes lacking a signal sequence were used to select export elements from Lactococcus lactis chromosomal DNA that could function as signal sequences. Fragments containing such elements were identified by their ability to direct the

  13. Environmental stress responses in Lactococcus lactis

    NARCIS (Netherlands)

    Sanders, JW; Venema, G; Kok, J

    1999-01-01

    Bacteria can encounter a variety of physical conditions during their life, Bacterial cells are able to survive these (often adverse) conditions by the induction of specific or general protection mechanisms. The lactic acid bacterium Lactococcus lactis is widely used for the production of cheese. Bef

  14. Heterologous Protein Expression by Lactococcus lactis

    NARCIS (Netherlands)

    Villatoro-Hernández, J.; Kuipers, O.P.; Saucedo-Cárdenas, O.; Montes-de-Oca-Luna, R.

    2012-01-01

    This chapter describes the use of Lactococcus lactis as a safe and efficient cell factory to produce heterologous proteins of medical interest. The relevance of the use of this lactic acid bacterium (LAB) is that it is a noncolonizing, nonpathogenic microorganism that can be delivered in vivo at a m

  15. Regulation of carbon catabolism in Lactococcus lactis.

    NARCIS (Netherlands)

    Aleksandrzak, T; Kowalczyk, M; Kok, J; Bardowski, J; Bielecki, S; Tramper, J; Polak, J

    2000-01-01

    The Lactococcus lactis IL1403 is a lactose negative, plasmid free strain. Nevertheless, it is able to hydrolyze lactose in the presence of cellobiose. In this work we describe identification of a gene involved in this process. The gene was found to be homologous to the sugar catabolism regulator, cc

  16. Functionality of Sortase A in Lactococcus lactis

    NARCIS (Netherlands)

    Dieye, Yakhya; Oxaran, Virginie; Ledue-Clier, Florence; Alkhalaf, Walid; Buist, Girbe; Juillard, Vincent; Lee, Chang Won; Piard, Jean-Christophe

    2010-01-01

    Lactococcus lactis IL1403 harbors a putative sortase A (SrtA) and 11 putative sortase substrates that carry the canonical LPXTG signature of such substrates. We report here on the functionality of SrtA to anchor five LPXTG substrates to the cell wall, thus suggesting that SrtA is the housekeeping so

  17. Procedure for quantifiable assessment of nutritional parameters influencing nisin production by Lactococcus lactis subsp. lactis.

    Science.gov (United States)

    Chandrapati, S; O'Sullivan, D J

    1998-08-27

    A modified rapid plate assay procedure was developed, that allowed quantifiable measurement of nisin production by Lactococcus lactis growing directly on agar media. Using this direct plate assay, several nutritional parameters were assessed for their influence on nisin production (as distinct from their influence on growth) by L. lactis subsp. lactis ATCC 11454 growing on standard M17 based media over 3 and 6 h incubation periods. Glucose was found to be the optimal carbon source tested, with glycerol having the greatest suppressive effect. The addition of salts suppressed nisin production on a per cell basis, except MnCl2. This direct plate method proved to be a good pilot assay for rapidly and quantifiably investigating the initial effects of different parameters on nisin production by L. lactis, prior to conducting more intensive broth batch culture assays. The data obtained in this study indicate that certain nutritional parameters can impose a repressive effect on nisin production. Elucidation of how these parameters control the amount of nisin produced will provide further insight into the regulation of nisin biosynthesis in L. lactis.

  18. Characterization of bacteriocins produced by Lactococcus lactis strains Caracterização de bacteriocinas produzidas por linhagens de Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    Izildinha Moreno

    2000-09-01

    Full Text Available Bacteriocins produced by fifteen strains of Lactococcus lactis (14 L. lactis subsp. lactis and one L. lactis subsp. cremoris were heat resistant, sensitive to several proteolytic enzymes and active over a wide range of pH. Their resistance to the heating was greatly influenced by the pH. Only the strain L. lactis subsp. lactis ITAL 383 produced a bacteriocin with a wide activity spectrum, similar to nisin of L. lactis subsp. lactis ATCC 11454. This bacteriocin inhibited closely related species and other Gram-positive microorganisms including Listeria monocytogenes and Staphylococcus aureus, but it was not active against the Gram-negative bacteria tested. The identification of partially purified antimicrobial compounds by SDS-PAGE showed that bacteriocin produced by strain ITAL 383 had the same molecular weight of nisin produced by L. lactis subsp. lactis ATCC 11454.Bacteriocinas resistentes ao aquecimento produzidas por quinze linhagens de Lactococcus lactis (14 L. lactis subsp. lactis e 1 L. lactis subsp. cremoris foram sensíveis à enzimas proteolíticas e ativas em uma ampla faixa de pH. A resistência dessas bacteriocinas ao aquecimento foi fortemente influenciada pelo pH do meio. Somente a linhagem L. lactis subsp. lactis ITAL 383 produziu uma bacteriocina com um amplo espectro de atividade, semelhante ao da nisina de L. lactis subsp. lactis ATCC 11454. Esta bacteriocina inibiu as espécies relacionadas e outros microorganismos gram-positivos, inclusive Listeria monocytogenes e Staphylococcus aureus, mas não as bactérias Gram-negativas examinadas. A identificação do composto antimicrobiano parcialmente purificado por SDS-PAGE revelou um peso molecular similar entre a bacteriocina ITAL 383 e a nisina de L. lactis subsp lactis ATCC 11454.

  19. Hypocholesterolaemic effect of yoghurt containing Bifidobacterium pseudocatenulatum G4 or Bifidobacterium longum BB536.

    Science.gov (United States)

    Al-Sheraji, Sadeq Hasan; Ismail, Amin; Manap, Mohd Yazid; Mustafa, Shuhaimi; Yusof, Rokiah Mohd; Hassan, Fouad Abdulrahman

    2012-11-15

    The effect of a yoghurt supplement containing Bifidobacterium pseudocatenulatum G4 or Bifidobacterium longum BB536 on plasma lipids, lipid peroxidation and the faecal excretion of bile acids was examined in rats fed a cholesterol-enriched diet. After 8 weeks, the rats in the positive control (PC) group who were fed the cholesterol-enriched diet showed significant increases in plasma total cholesterol (TC), low-density lipoprotein (LDL) cholesterol, and malondialdehyde (MDA). However, groups fed a cholesterol-enriched diet supplemented with yoghurt containing B. pseudocatenulatum G4 or B. longum BB536 had significantly lower plasma TC, LDL-C, very-low-density lipoprotein (VLDL) cholesterol, and MDA than had the PC group after 8 weeks of treatment. In addition, faecal excretion of bile acids was markedly increased in the rats fed the yoghurt containing B. pseudocatenulatum G4 or B. longum BB536 as compared to the PC and NC groups.

  20. Bacteriocinogenic Lactococcus lactis subsp. lactis DF04Mi isolated from goat milk: evaluation of the probiotic potential.

    Science.gov (United States)

    Furtado, Danielle N; Todorov, Svetoslav D; Landgraf, Mariza; Destro, Maria T; Franco, Bernadette D G M

    2014-01-01

    Lactic acid bacteria capable of producing bacteriocins and presenting probiotic potential open innovative technological applications in the dairy industry. In this study, a bacteriocinogenic strain (Lactococcus lactis subsp. lactis DF4Mi) was isolated from goat milk, and studied for its probiotic potential. Lc. lactis DF4Mi was resistant to acidic pH and oxbile, presented co-aggregation with Listeria monocytogenes, and was not affected by several drugs from different generic groups, being sensitive to most tested antibiotics. These properties indicate that this Lc. lactis strain can be used for enhancement of dairy foods safety and quality, in combination with potential probiotic properties.

  1. Bacteriocinogenic Lactococcus lactis subsp: lactis DF04Mi isolated from goat milk: Evaluation of the probiotic potential

    Directory of Open Access Journals (Sweden)

    Danielle N. Furtado

    2014-09-01

    Full Text Available Lactic acid bacteria capable of producing bacteriocins and presenting probiotic potential open innovative technological applications in the dairy industry. In this study, a bacteriocinogenic strain (Lactococcus lactis subsp. lactis DF4Mi was isolated from goat milk, and studied for its probiotic potential. Lc. lactis DF4Mi was resistant to acidic pH and oxbile, presented co-aggregation with Listeria monocytogenes, and was not affected by several drugs from different generic groups, being sensitive to most tested antibiotics. These properties indicate that this Lc. lactis strain can be used for enhancement of dairy foods safety and quality, in combination with potential probiotic properties.

  2. Comparative genomics of bifidobacterium, lactobacillus and related probiotic genera

    DEFF Research Database (Denmark)

    Lukjancenko, Oksana; Ussery, David; Wassenaar, Trudy M.

    2012-01-01

    Six bacterial genera containing species commonly used as probiotics for human consumption or starter cultures for food fermentation were compared and contrasted, based on publicly available complete genome sequences. The analysis included 19 Bifidobacterium genomes, 21 Lactobacillus genomes, 4...

  3. Bile resistance mechanisms in Lactobacillus and Bifidobacterium

    Directory of Open Access Journals (Sweden)

    Lorena eRuiz

    2013-12-01

    Full Text Available Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. Most of the probiotic bacteria currently available in the market belong to the genera Lactobacillus and Bifidobacterium, and specific health-promoting activities, such as treatment of diarrhea or amelioration of gastrointestinal discomfort, have been attributed to them. In order to be able to survive the gastrointestinal transit and transiently colonise our gut, these bacteria must be able to counteract the deleterious action of bile salts, which are the main components of bile. Bile salts are detergent-like biological substances synthesised in the liver from cholesterol. Host enzymes conjugate the newly synthesised free bile acids in the liver with the amino acids glycine or taurine, generating conjugated bile salts. These compounds are stored in the gall bladder and they are released into the duodenum during digestion to perform their physiological function, which is the solubilisation of fat coming from diet. These bile salts possess strong antimicrobial activity, since they are able to disorganize the structure of the cell membrane, as well as trigger DNA damage. This means that bacteria inhabiting our intestinal tract must have intrinsic resistance mechanisms to cope with bile salts. To do that, Lactobacillus and Bifidobacterium display a variety of proteins devoted to the efflux of bile salts or protons, to modify sugar metabolism or to prevent protein misfolding. In this manuscript, we review and discuss specific bile resistance mechanisms, as well as the processes responsible for the adaptation of bifidobacteria and lactobacilli to bile.

  4. Alternative lactose catabolic pathway in Lactococcus lactis IL1403

    NARCIS (Netherlands)

    Aleksandrzak-Piekarczyk, T; Kok, J; Renault, P; Bardowski, J

    2005-01-01

    In this study, we present a glimpse of the diversity of Lactococcus lactis subsp. lactis IL1403 beta-galactosidase phenotype-negative mutants isolated by negative selection on solid media containing cellobiose or lactose and X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside), and we identif

  5. Transforming Lactococcus lactis into a microbial cell factory

    DEFF Research Database (Denmark)

    Petersen, Kia Vest

    . To simplify further analysis arcA encoding the arginine deiminase was deleted, thus eliminating the arginine catabolism. We found that in L. lactis KF147 xylose is metabolized through two pathways namely the phosphoketolase pathway and the non-oxidative part of the pentose phosphate pathway. The only products...... the potential of Lactococcus lactis as a platform organism for production of biofuels and-chemicals with a focus on characterization and optimization of the xylose metabolism. The plant isolate L. lactis KF147 was selected as the potential platform organism due to its natural ability to utilize both the pentose...... for the next round of integration. The xylose metabolism in L. lactis KF147 was characterized in a defined medium supplemented with 0.2%, 1% or 3% (w/v) xylose. The defined medium contains free arginine, and it was found that L. lactis KF147 co-metabolizes the arginine through the arginine deiminase pathway...

  6. Heterologous Gene Expression in Lactococcus lactis subsp. lactis : Synthesis, Secretion, and Processing of the Bacillus subtilis Neutral Protease

    NARCIS (Netherlands)

    Guchte, Maarten van de; Kodde, Jan; Vossen, Jos M.B.M. van der; Kok, Jan; Venema, Gerard

    1990-01-01

    The Bacillus subtilis nprE gene lacking its own promoter sequence was inserted in the lactococcal expression vector pMG36e. Upon introduction of the recombinant plasmid into Lactococcus lactis subsp. lactis strain MG1363, neutral protease activity could be visualized by the appearance of large clear

  7. Suitability of Lactococcus lactis subsp lactis ATCC 11454 as a protective culture for lightly preserved fish products

    DEFF Research Database (Denmark)

    Wessels, Stephen Wallace; Huss, Hans Henrik

    1996-01-01

    This study is part of strategy to control the human pathogen Listeria monocytogenes in lightly preserved fish products by using food-grade lactic acid bacteria. When the nisin-producing Lactococcus lactis subsp lactis ATCC 11454 was cultured in the same vessel as L-monocytogenes Scott A in brain...

  8. Detection and viability of Lactococcus lactis throughout cheese ripening.

    Science.gov (United States)

    Ruggirello, Marianna; Dolci, Paola; Cocolin, Luca

    2014-01-01

    Recent evidences highlighted the presence of Lactococcus lactis during late cheese ripening. For this reason, the role of this microorganism, well known as dairy starter, should be reconsidered throughout cheese manufacturing and ripening. Thus, the main objective of this study was to develop a RT-qPCR protocol for the detection, quantification and determination of the viability of L. lactis in ripened cheese samples by direct analysis of microbial nucleic acids. Standard curves were constructed for the specific quantification of L. lactis in cheese matrices and good results in terms of selectivity, correlation coefficient and efficiency were obtained. Thirty-three ripened cheeses were analyzed and, on the basis of RNA analysis, twelve samples showed 106 to 108 CFU of L. lactis per gram of product, thirteen from 103 to 105 CFU/g, and in eight cheeses, L. lactis was not detected. Traditional plating on M17 medium led to loads ranging from 105 to 109 CFU/g, including the cheese samples where no L. lactis was found by RT-qPCR. From these cheeses, none of the colonies isolated on M17 medium was identified as L. lactis species. These data could be interpreted as a lack of selectivity of M17 medium where colony growth is not always related to lactococcal species. At the same time, the absence or low abundance of L. lactis isolates on M17 medium from cheese where L. lactis was detected by RT-qPCR support the hypothesis that L. lactis starter populations are mainly present in viable but not culturable state during ripening and, for this reason, culture-dependent methods have to be supplemented with direct analysis of cheese.

  9. Detection and viability of Lactococcus lactis throughout cheese ripening.

    Directory of Open Access Journals (Sweden)

    Marianna Ruggirello

    Full Text Available Recent evidences highlighted the presence of Lactococcus lactis during late cheese ripening. For this reason, the role of this microorganism, well known as dairy starter, should be reconsidered throughout cheese manufacturing and ripening. Thus, the main objective of this study was to develop a RT-qPCR protocol for the detection, quantification and determination of the viability of L. lactis in ripened cheese samples by direct analysis of microbial nucleic acids. Standard curves were constructed for the specific quantification of L. lactis in cheese matrices and good results in terms of selectivity, correlation coefficient and efficiency were obtained. Thirty-three ripened cheeses were analyzed and, on the basis of RNA analysis, twelve samples showed 106 to 108 CFU of L. lactis per gram of product, thirteen from 103 to 105 CFU/g, and in eight cheeses, L. lactis was not detected. Traditional plating on M17 medium led to loads ranging from 105 to 109 CFU/g, including the cheese samples where no L. lactis was found by RT-qPCR. From these cheeses, none of the colonies isolated on M17 medium was identified as L. lactis species. These data could be interpreted as a lack of selectivity of M17 medium where colony growth is not always related to lactococcal species. At the same time, the absence or low abundance of L. lactis isolates on M17 medium from cheese where L. lactis was detected by RT-qPCR support the hypothesis that L. lactis starter populations are mainly present in viable but not culturable state during ripening and, for this reason, culture-dependent methods have to be supplemented with direct analysis of cheese.

  10. The effect of lactose, NaCl and an aero/anaerobic environment on the tyrosine decarboxylase activity of Lactococcus lactis subsp. cremoris and Lactococcus lactis subsp. lactis.

    Science.gov (United States)

    Buňková, Leona; Buňka, František; Pollaková, Eva; Podešvová, Tereza; Dráb, Vladimír

    2011-05-27

    The aim of this work was to study, under model conditions, combined effects of the concentration of lactose (0-1% w/v), NaCl (0-2% w/v) and aero/anaerobiosis on the growth and tyramine production in 3 strains of Lactococcus lactis subsp. lactis and 2 strains of L. lactis subsp. cremoris. The levels of the factors tested were chosen with respect to the conditions which can occur during the real process of natural cheese production, including the culture temperature (10 ± 1°C). In all strains tested, tyrosine decarboxylation was most influenced by NaCl concentration; the highest production of tyramine was obtained within the culture with the highest (2% w/v) salt concentration applied. Two of the strains L. lactis subsp. lactis produced tyramine only in broth with the highest NaCl concentration tested. In the remaining 3 strains of L. lactis, tyramine was detected under all conditions applied. The tested concentration of lactose and aero/anaerobiosis had a less significant effect on tyramine decarboxylation. However, it was also found that at the same concentrations of NaCl and lactose, a higher amount of tyramine was detected under anaerobic conditions. In all strains tested, tyramine decarboxylation started during the active growth phase of the cells.

  11. [Surface proteins of bacteria of the genus Bifidobacterium].

    Science.gov (United States)

    Dylus, Ewa; Buda, Barbara; Górska-Frączek, Sabina; Brzozowska, Ewa; Gamian, Andrzej

    2013-05-13

    Beneficial effects due to the presence of probiotic bacteria of the genus Bifidobacterium in the human intestinal tract are still an interesting object of study. So far activities have been confirmed of bifidobacteria in stimulation of the host immune system, stimulation of tumor cell apoptosis, improvement of bowel motility, alleviation of symptoms of lactose intolerance, cholesterol lowering capacity, prevention and treatment of diarrhea and irritable bowel syndrome, alleviation of allergy or atopic dermatitis, maintenance of homeostasis of the intestine, and stimulation of the development of normal intestinal microflora in infants. A multitude of therapeutic properties encourages researchers to investigate the possibility of using the potential of Bifidobacterium in the prevention and treatment of other conditions such as rheumatoid arthritis and depression. Although it is known that the beneficial effects are due to intestinal mucosal colonization by these bacteria, the cell components responsible for the colonization are still not determined. In addition to the beneficial effects of probiotic administration, there were also negative effects including sepsis. Therefore research has been directed to identify specific components of Bifidobacterium responsible for probiotic effects. Currently researchers are focused on identifying, isolating and evaluating the properties of surface proteins that are probably involved in the adhesion of bacterial cells to the intestinal epithelium, improving colonization. This paper is an overview of current knowledge on Bifidobacterium surface proteins. The ways of transport and anchoring proteins in Gram-positive bacterial cells, the assembly of cell wall, and a description of the genus Bifidobacterium are presented.

  12. Microbes of fermented kefir-like using combination of kefir grains and Bifidobacterium longum

    Directory of Open Access Journals (Sweden)

    Sri Usmiati

    2005-03-01

    Full Text Available The objectives of research were to find out physico-chemical characters and to detect flavor volatile compound of kefir-like. Material used was skim milk TS 9.5% which was heated at 85oC for 30 minutes and cooled at 22oC before innoculation of the starter. Microorganisms used were (a Lactobacillus acidophilus P155110, (b Lactobacillus delbrueckii subsp. Bulgaricus NCIMB 11778, (c Lactococcus lactis P155610, (d Leuconostoc mesenteroides subsp. dextranicum NCIMB 3350, (e Acetobacter aceti P154810, (f Bifidobacterium longum BF1, and (g Saccharomyces cerevisiae P156252. The treatments consist of P1 = without (b; P2 = without (a; and P3= used (a until (g. The physico-chemical characters identified were lactic acid and lactose percentages, pH, viscosity, organoleptic test for intensity of kefir-like sensory attributes. Results indicated that B. longum was potential bacterium use for starter combination on kefir-like making. The use starter P1 combination has high acidity and viscosity, low pH and lactose percentage, and high intensity on attribute creamy-white color, soft and curdle consistency, and kefir specific aroma on kefir-like. Volatile compound acid group were dominate by high acidity character on kefir-like resulted from starter P1 combination. Compound of 3-hydroxi-2-butanone (acetoin was affecting butter-like of P3 character. This compound resulted from which is a character of fermented milk flavor was not detected on P1 kefir-like.

  13. POTENTIAL OF Lactococcus lactis subsp. lactis MTCC 3041 AS A BIOPRESERVATIVE

    Directory of Open Access Journals (Sweden)

    Neha Sharma

    2013-10-01

    Full Text Available Lactic acid bacteria especially in developing countries can be exploited against frequently occurring spoilage organisms of fresh fruits and vegetables in addition to pathogens. Keeping in views this antagonism imparted by bacteria Lactococci, the present study was taken and effectiveness of bacteriocin of Lactococci was also studied in preservatives and enzymes. Lactic acid bacteria Lactococcus lactis subs. Lactis MTCC 3041 was used as bacteriocin producer strain. Isolation of most frequently occurring spoilage organisms from spoiled Mango and Kinnow was done by microbiological procedures and were identified by microscopic studies as Isolate 1 and Isolate 2. It has limited use in processed salted food as no zone of inhibition was observed at and above 5% NaCl (w/v.0.3% (w/v is the minimum concentration of KMS that provides stress to the microorganism for the production of bacteriocin. It is not suitable for food having sodium benzoate as preservative as with increase in concentration growth of Lactococcus lactis decreases. Presence of bacteriocin hinders the growth of the isolate 1 as fresh weight of the mycelium in test sample is 7.09% less than the control. Being non-pathogenic this organism can be safely used against spoilage organisms in addition to food borne pathogens.

  14. EFEITO DE PROTETORES E TRATAMENTOS DE ESTRESSE NA SOBREVIVÊNCIA DE LACTOCOCCUS LACTIS SUBSP LACTIS AO CONGELAMENTO Effect of protective and stress treatment on survival of Lactococcus lactis subsp. lactis to freezing

    Directory of Open Access Journals (Sweden)

    Eliana dos Santos Leandro

    2013-02-01

    Full Text Available O efeito de substâncias crioprotetoras e de tratamentos subletais de estresse foi avaliado no aumento da tolerância ao congelamento em Lactococcus lactis subsp lactis PD 6.9. Todas as substâncias crioprotetoras avaliadas aumentaram a sobrevivência de L. lactis subsp lactis PD 6.9 a estocagem de 15 dias a - 20 ºC. Entretanto, o leite desnatado reconstituído a 10 % foi o que conferiu maior proteção. Quanto à exposição da suspensão de células a tratamentos subletais de estresse, a exposição a 10 ºC por 4 horas foi capaz de manter a sobrevivência de L. lactis subsp lactis PD 6.9 estável durante 70 dias de estocagem a - 20 ºC. O tratamento a 40 ºC por 30 minutos conferiu proteção durante 70 dias de estocagem a - 20 ºC quando comparado com a suspensão de células que não recebeu nenhum tratamento antes do congelamento. A aplicação desses tratamentos é importante para assegurar a viabilidade de L. lactis subsp lactis PD 6.9 no decorrer do período de estocagem, e também de assegurar a qualidade sensorial e microbiológica de alimentos obtidos por processos de fermentação.

  15. Chorioamnionitis due to Lactococcus lactis cremoris: A case report

    Directory of Open Access Journals (Sweden)

    F. Azouzi

    2015-07-01

    Full Text Available Lactococcus lactis cremoris is rarely involved in human pathology. A thirty two-year old pregnant woman with premature rupture of membrane history presented with chorioamnionitis due to L. lactis cremoris. She underwent an emergency caesarian section and was treated with antibiotics including the association of amoxicillin and clavulanic acid. She was completely recovered. This is the first case to our knowledge of chorioamnionitis due to this organism.

  16. Chorioamnionitis due to Lactococcus lactis cremoris: A case report

    OpenAIRE

    F. Azouzi; C. Chahed; Marzouk, M.; A. Ferjani; N. Hannechi; M. Fekih; Y. Ben Salem; J. Boukadida

    2015-01-01

    Lactococcus lactis cremoris is rarely involved in human pathology. A thirty two-year old pregnant woman with premature rupture of membrane history presented with chorioamnionitis due to L. lactis cremoris. She underwent an emergency caesarian section and was treated with antibiotics including the association of amoxicillin and clavulanic acid. She was completely recovered. This is the first case to our knowledge of chorioamnionitis due to this organism.

  17. Expression of biologically active murine interleukin-18 in Lactococcus lactis.

    Science.gov (United States)

    Feizollahzadeh, Sadegh; Khanahmad, Hossein; Rahimmanesh, Ilnaz; Ganjalikhani-Hakemi, Mazdak; Andalib, Alireza; Sanei, Mohammad Hossein; Rezaei, Abbas

    2016-11-01

    The food-grade bacterium Lactococcus lactis is increasingly used for heterologous protein expression in therapeutic and industrial applications. The ability of L. lactis to secrete biologically active cytokines may be used for the generation of therapeutic cytokines. Interleukin (IL)-18 enhances the immune response, especially on mucosal surfaces, emphasizing its therapeutic potential. However, it is produced as an inactive precursor and has to be enzymatically cleaved for maturation. We genetically manipulated L. lactis to secrete murine IL-18. The mature murine IL-18 gene was inserted downstream of a nisin promoter in pNZ8149 plasmid and the construct was used to transform L. lactis NZ3900. The transformants were selected on Elliker agar and confirmed by restriction enzyme digestion and sequencing. The expression and secretion of IL-18 protein was verified by SDS-PAGE, western blotting and ELISA. The biological activity of recombinant IL-18 was determined by its ability to induce interferon (IFN)-γ production in L. lactis co-cultured with murine splenic T cells. The amounts of IL-18 in bacterial lysates and supernatants were 3-4 μg mL(-1) and 0.6-0.7 ng mL(-1), respectively. The successfully generated L. lactis strain that expressed biologically active murine IL-18 can be used to evaluate the possible therapeutic effects of IL-18 on mucosal surfaces.

  18. Detection and characterization of bacteriocin-producing Lactococcus lactis strains Detecção e caracterização de Lactococcus lactis produtores de bacteriocinas

    Directory of Open Access Journals (Sweden)

    Izildinha Moreno

    1999-04-01

    Full Text Available One hundred sixty seven strains of Lactococcus lactis were screened for bacteriocin production by well diffusion assay of GM17 agar. Fourteen (8.4% produced antimicrobial activity other than organic acids, bacteriophages or hydrogen peroxide. The frequency of bacteriocin production ranged from 2% in L. lactis subsp. cremoris up to 12% in L. lactis subsp. lactis. Antimicrobial activities were not observed in any strain of L. lactis subsp. lactis var. diacetylactis. Among thirteen bacteriocin-producing strains and two nisin-producing strains (L. lactis subsp. lactis ATCC 11454 and L. lactis subsp. lactis CNRZ 150, eight (53% were characterized as lactose-positive (Lac+ and proteinase-negative (Prt-. The bacteriocin-producing cultures were also characterized on the basis of plasmid content. All strains had 2 to 7 plasmids with molecular weights varying from 0.5 to 28.1 Mdal. Four strains (ITAL 435, ITAL 436, ITAL 437 and ITAL 438 showed identical profiles and the other were quite distinct.Um total de 167 linhagens de L. lactis foi selecionado para os testes de produção de bacteriocinas pelo método de difusão em poços em agar GM17. Desse total, 14 (8.4% produziram substâncias inibidoras que não foram associadas com ácidos orgânicos, peróxido de hidrogênio e bacteriófagos. A frequência de produção de bacteriocinas variou de 2% em L. lactis subsp. cremoris a 12% em L. lactis subsp. lactis. Nenhuma das linhagens de L. lactis subsp. lactis var. diacetylactis produziu substâncias inibidoras. De 13 linhagens produtoras de bacteriocinas e duas de nisina (L. lactis subsp. lactis ATCC 11454 e L. lactis subsp. lactis CNRZ 150, 8 (53% foram caracterizadas como lactose-positivas (Lac+ e proteinase-negativas (Prt-. As linhagens produtoras de bacteriocinas também foram caracterizadas no seu conteúdo de plasmídios. Elas apresentaram de 2 a 7 plasmídios, com pesos moleculares aproximados de 0.5 a 28.1 Mdal. Quatro linhagens (ITAL 435, ITAL 436

  19. Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis

    DEFF Research Database (Denmark)

    Sprogøe, Desiree; van den Broek, Lambertus A M; Mirza, Osman

    2004-01-01

    phosphorylase from Bifidobacterium adolescentis (BiSP) refined at 1.77 A resolution. It represents the first 3D structure of a sucrose phosphorylase and is the first structure of a phosphate-dependent enzyme from the glycoside hydrolase family 13. The structure of BiSP is composed of the four domains A, B, B...

  20. Complete genome sequence of Bifidobacterium bifidum S17.

    NARCIS (Netherlands)

    Zhurina, D.; Zomer, A.L.; Gleinser, M.; Brancaccio, V.F.; Auchter, M.; Waidmann, M.S.; Westermann, C.; Sinderen, D. van; Riedel, C.U.

    2011-01-01

    Here, we report on the first completely annotated genome sequence of a Bifidobacterium bifidum strain. B. bifidum S17, isolated from feces of a breast-fed infant, was shown to strongly adhere to intestinal epithelial cells and has potent anti-inflammatory activity in vitro and in vivo. The genome se

  1. Genomics and ecological overview of the genus Bifidobacterium.

    Science.gov (United States)

    Turroni, Francesca; van Sinderen, Douwe; Ventura, Marco

    2011-09-01

    Members of the genus Bifidobacterium are high G+C Gram positive bacteria belonging to the phylum Actinobacteria, and represent common inhabitants of the gastro-intestinal tract (GIT) of mammals, birds and certain cold-blooded animals. The overall microbial population that resides in the GIT, referred to as the "gut microbiota", is an extremely complex community of microorganisms whose functions are believed to have a significant impact on human physiology. Different ecological relationships between bifidobacteria and their host can be developed, ranging from opportunistic pathogenic interactions (e.g. in the case of Bifidobacterium dentium) to a commensal or even health-promoting relationship (e.g. in the case of Bifidobacterium bifidum and Bifidobacterium breve species). Among the known health-promoting or probiotic microorganisms, bifidobacteria represent one of the most dominant group and some bifidobacterial species are frequently used as the probiotic ingredient in many functional foods. However, despite the generally accepted importance of bifidobacteria as constituents of the human microbiota, there is only limited information available on their phylogeny, physiology and genetics. Moreover, host-microbiota interactions and cross-talk between different members of the gut microbiota are far from completely understood although they represent a crucial factor in the development and maintenance of human physiology and immune system. The aim of this review is to highlight the genetic and functional features of bifidobacteria residing in the human GIT using genomic and ecology-based information.

  2. Glycosyl hydrolases from Bifidobacterium adolescentis DSM20083. An overview

    NARCIS (Netherlands)

    Broek, van den L.A.M.; Hinz, S.W.A.; Beldman, G.; Doeswijk-Voragen, C.H.L.; Vincken, J.P.; Voragen, A.G.J.

    2005-01-01

    It is claimed that bifidobacteria have several health-promoting effects. To increase the amount of bifidobacteria in the colon the concept of probiotics and/or prebiotics can be applied. Bifidobacterium adolescentis is one of the main species of bifidobacteria in the gastro-intestinal tract of human

  3. Surface proteins of bacteria of the genus Bifidobacterium 

    Directory of Open Access Journals (Sweden)

    Ewa Dylus

    2013-05-01

    Full Text Available Beneficial effects due to the presence of probiotic bacteria of the genus Bifidobacterium in the human intestinal tract are still an interesting object of study. So far activities have been confirmed of bifidobacteria in stimulation of the host immune system, stimulation of tumor cell apoptosis, improvement of bowel motility, alleviation of symptoms of lactose intolerance, cholesterol lowering capacity, prevention and treatment of diarrhea and irritable bowel syndrome, alleviation of allergy or atopic dermatitis, maintenance of homeostasis of the intestine, and stimulation of the development of normal intestinal microflora in infants. A multitude of therapeutic properties encourages researchers to investigate the possibility of using the potential of Bifidobacterium in the prevention and treatment of other conditions such as rheumatoid arthritis and depression. Although it is known that the beneficial effects are due to intestinal mucosal colonization by these bacteria, the cell components responsible for the colonization are still not determined. In addition to the beneficial effects of probiotic administration, there were also negative effects including sepsis. Therefore research has been directed to identify specific components of Bifidobacterium responsible for probiotic effects. Currently researchers are focused on identifying, isolating and evaluating the properties of surface proteins that are probably involved in the adhesion of bacterial cells to the intestinal epithelium, improving colonization. This paper is an overview of current knowledge on Bifidobacterium surface proteins. The ways of transport and anchoring proteins in Gram-positive bacterial cells, the assembly of cell wall, and a description of the genus Bifidobacterium are presented.

  4. [Characteristics and identification of bacteriocins produced by Lactococcus lactis subsp. lactis 194-K].

    Science.gov (United States)

    Ustiugova, E A; Timofeeva, A V; Stoianova, L G; Netrusov, A I; Katrukha, G S

    2012-01-01

    The Lactococcus lactis subsp. lactis 194-K strain has been established to be able to produce two bacteriocins, one of which was identified as the known lantibiotic nisin A, and the other 194-D bacteriocin represents a polypeptide with a 2589-Da molecular mass and comprises 20 amino acid residues. Both bacteriocins were produced in varying proportions in all of the studied nutrient media, which support the growth of the producer. Depending on the cultivation medium, the nisin A content was 380- to 1123-fold lower in the 194-K stain culture fluid than that of the 194-D peptide. In comparision to to nisin A Bacteriocin 194-D possessed a wide range of antibacterial activity and suppressed the growth of both Gram-positive and Gram-negative bacteria. An optimal medium for 194-D bacteriocin synthesis was shown to be a fermentation medium which contained yeast extract, casein hydrolysate, and potassium phosphate. The biosynthesis ofbacteriocin 194-D by the 194-K strain in these media occurred parallel to producer growth, and its maximal accumulation in the culture fluid was observed at 14-20 h of the strain's growth.

  5. LANTIBIOTIC NISIN: NATURAL PRESERVATIVE FROM LACTOCOCCUS LACTIS

    Directory of Open Access Journals (Sweden)

    Suganthi.V

    2012-01-01

    Full Text Available The increasing demand for high quality safe foods that are not extensively processed has created a niche for natural food preservative. Studies confirm that food allergies due to chemical preservatives affect as much as 2.5% of the population. Recent research had suggested bacteriocins (Nisin are the ideal biological food preservative. Nisin was proteinaceous antibacterial substances produced by Lactococcus lactis, a homofermentative bacterium. Naturally nisin occurs in two different forms nisin A and nisin Z. Nisin has wide range of inhibitory mode of action on Gram negative bacteria and food borne pathogens. Food preservation is a continuous war against the microorganisms spoiling the food or making it unsafe. So, nisin is actually the only lantibiotic bacteriocins used as a food preservative. This review paper will discuss about the Lactococcal strain used for the production of nisin, different forms of nisin, the mode of action of nisin, the cost reductive methods for the production and purification of nisin. So that it can be used in large scale industry for the high yield of nisin and the wide application of nisin in food industries.

  6. Investigation of glycerol assimilation and cofactor metabolism in Lactococcus lactis

    DEFF Research Database (Denmark)

    Holm, Anders Koefoed

    glyceraldehyde-3-phophate dehydrogenase and the pyruvate dehydrogenase complex, disrupting flow through the central metabolism and ATP production. If this is the case, the question remains, as to why the excess redox is not simply removed by respiration. The results from this investigation have provided...... cycle assessment of the GLYFINERY project, screening of L. lactis spp. for glycerol utilization, engineering of glycerol metabolism in L. lactis and finally an investigation into perturbation of energy metabolism in L. lactis. The work from the life cycle assessment resulted in two reports, detailing......: anaerobic, aerobic and respiration permissive growth in combination with either glycerol as a sole substrate or with co-metabolization of glycerol with common sugar substrates. Although no growth on glycerol was seen, both positive and detrimental effects were observed from cultures with glycerol...

  7. Genomic encyclopedia of type strains of the genus Bifidobacterium.

    Science.gov (United States)

    Milani, Christian; Lugli, Gabriele Andrea; Duranti, Sabrina; Turroni, Francesca; Bottacini, Francesca; Mangifesta, Marta; Sanchez, Borja; Viappiani, Alice; Mancabelli, Leonardo; Taminiau, Bernard; Delcenserie, Véronique; Barrangou, Rodolphe; Margolles, Abelardo; van Sinderen, Douwe; Ventura, Marco

    2014-10-01

    Bifidobacteria represent one of the dominant microbial groups that are present in the gut of various animals, being particularly prevalent during the suckling stage of life of humans and other mammals. However, the overall genome structure of this group of microorganisms remains largely unexplored. Here, we sequenced the genomes of 42 representative (sub)species across the Bifidobacterium genus and used this information to explore the overall genetic picture of this bacterial group. Furthermore, the genomic data described here were used to reconstruct the evolutionary development of the Bifidobacterium genus. This reconstruction suggests that its evolution was substantially influenced by genetic adaptations to obtain access to glycans, thereby representing a common and potent evolutionary force in shaping bifidobacterial genomes.

  8. Estudo dos parâmetros da ultrafiltração de permeado de soro de queijo fermentado por Lactococcus lactis subsp. lactis Ultrafiltration conditions of whey permeate fermented by Lactococcus lactis subsp. lactis

    Directory of Open Access Journals (Sweden)

    Viviane BRONSTEIN

    1998-04-01

    Full Text Available Permeado de soro doce, suplementado com extrato de levedura e peptona, foi utilizado como meio de crescimento para Lactococcus lactis subsp. lactis. No final da fase exponencial de crescimento, o meio de cultura fermentado foi submetido a uma ultrafiltração com o objetivo de concentrar o microrganismo. Foram realizados 6 processamentos diferentes, nos quais variou-se as condições iniciais da ultrafiltração, tendo sido avaliados os seguintes parâmetros: porosidade da membrana, pH e número de células viáveis no permeado e no retentado, a fim de ser estudado a influência de cada parâmetro na taxa de permeação da ultrafiltração. As membranas utilizadas foram eficazes como meio de barragem para o microrganismo Lactococcus lactis subsp. lactis, ficando o retentado com uma média celular de 10(8 ufc/ml e o permeado com uma média celular de 10² ufc/ml. Membranas de diferentes porosidades tiveram taxas de fluxo semelhantes. O aumento da concentração celular provocou a diminuição do fluxo. O pH também influenciou a taxa de permeação, havendo um aumento do fluxo quando foi utilizado um pH inicial mais alto.Cheese whey permeate supplemented with yeast extract and peptone was used as a growth medium for the bacteria Lactococcus lactis subsp. lactis. At the end of the exponential growth phase, the fermented growth medium was ultrafiltered to concentrate the microorganism and to evaluate the effect of the membrane porosity, inicial UF pH and cellular concentration in permeation rate during the ultrafiltration process. The membranes used were efficient as a mean of a barrage for the Lactococcus lactis subsp. lactis. On average, the cellular concentrations were 10(8 CFU/mL and 10² CFU/mL for retentate and permeate, respectively. Membranes of different porosities had very similar flux rates. Better flow rates were obtained with inicial UF pH 6,5 and with the minors micrrorganism concentration.

  9. Cloning and expression of a novel lactococcal aggregation factor from Lactococcus lactis subsp. lactis BGKP1

    Directory of Open Access Journals (Sweden)

    Kojic Milan

    2011-12-01

    Full Text Available Abstract Background Aggregation may play a main role in the adhesion of bacteria to the gastrointestinal epithelium and their colonization ability, as well as in probiotic effects through co-aggregation with intestinal pathogens and their subsequent removal. The aggregation phenomenon in lactococci is directly associated with the sex factor and lactose plasmid co-integration event or duplication of the cell wall spanning (CWS domain of PrtP proteinase. Results Lactococcus lactis subsp. lactis BGKP1 was isolated from artisanal semi-hard homemade cheese and selected due to its strong auto-aggregation phenotype. Subsequently, non-aggregating derivative (Agg- of BGKP1, designated as BGKP1-20, was isolated, too. Comparative analysis of cell surface proteins of BGKP1 and derivative BGKP1-20 revealed a protein of approximately 200 kDa only in the parental strain BGKP1. The gene involved in aggregation (aggL was mapped on plasmid pKP1 (16.2 kb, cloned and expressed in homologous and heterologous lactococci and enterococci. This novel lactococcal aggregation protein was shown to be sufficient for cell aggregation in all tested hosts. In addition to the aggL gene, six more ORFs involved in replication (repB and repX, restriction and modification (hsdS, transposition (tnp and possible interaction with mucin (mbpL were also located on plasmid pKP1. Conclusion AggL is a new protein belonging to the collagen-binding superfamily of proteins and is sufficient for cell aggregation in lactococci.

  10. Luciferase detection during stationary phase in Lactococcus lactis

    NARCIS (Netherlands)

    Bachmann, H.; Santos, dos F.; Kleerebezem, M.; Hylckama Vlieg, van J.E.T.

    2007-01-01

    The luminescence signal of luxAB-encoded bacterial luciferase strongly depends on the metabolic state of the host cell, which restricts the use of this reporter system to metabolically active bacteria. Here we show that in stationary-phase cells of Lactococcus lactis, detection of luciferase is sign

  11. Nucleotide metabolism in Lactococcus lactis: Salvage pathways of exogenous pyrimidines

    DEFF Research Database (Denmark)

    Martinussen, Jan; Andersen, Paal Skytt; Hammer, Karin

    1994-01-01

    By measuring enzyme activities in crude extracts and studying the effect of toxic analogs (5-fluoropyrimidines) on cell growth, the metabolism of pyrimidines in Lactococcus lactis was analyzed. Pathways by which uracil, uridine, deoxyuridine, cytidine, and deoxycytidine are metabolized in L. lact...

  12. Regulatory phenotyping reveals important diversity within the species Lactococcus lactis

    NARCIS (Netherlands)

    Bachmann, H.; Starrenburg, M.; Dijkstra, A.; Molenaar, D.; Kleerebezem, M.; Rademaker, J.L.W.; Hylckama Vlieg, van J.E.T.

    2009-01-01

    The diversity in regulatory phenotypes among a collection of 84 Lactococcus lactis strains isolated from dairy and nondairy origin was explored. The specific activities of five enzymes were assessed in cell extracts of all strains grown in two different media, a nutritionally rich broth and a relati

  13. Transcriptional regulation of central amino acid metabolism in Lactococcus lactis

    NARCIS (Netherlands)

    Larsen, Rasmus

    2005-01-01

    This thesis describes the functional characterisation of the transcriptional regulators GlnR, ArgR and AhrC of Lactococcus lactis, which are responsible for the control of genes involved in the metabolism of the amino acids glutamine, glutamate and arginine. A chromosomal glnR deletion mutant was ma

  14. A review on Lactococcus lactis: from food to factory.

    Science.gov (United States)

    Song, Adelene Ai-Lian; In, Lionel L A; Lim, Swee Hua Erin; Rahim, Raha Abdul

    2017-04-04

    Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features of this gram-positive lactic acid non-colonizing gut bacteria include its generally recognized as safe (GRAS) status, probiotic properties, the absence of inclusion bodies and endotoxins, surface display and extracellular secretion technology, and a diverse selection of cloning and inducible expression vectors. This have made L. lactis a desirable and promising host on par with other well established model bacterial or yeast systems such as Escherichia coli, Salmonella cerevisiae and Bacillus subtilis. In this article, we review recent technological advancements, challenges, future prospects and current diversified examples on the use of L. lactis as a microbial cell factory. Additionally, we will also highlight latest medical-based applications involving whole-cell L. lactis as a live delivery vector for the administration of therapeutics against both communicable and non-communicable diseases.

  15. Nisin-Producing Lactococcus lactis Strains Isolated from Human Milk

    OpenAIRE

    Beasley, Shea S.; Saris, Per E.J.

    2004-01-01

    Characterization by partial 16S rRNA gene sequencing, ribotyping, and green fluorescent protein-based nisin bioassay revealed that 6 of 20 human milk samples contained nisin-producing Lactococcus lactis bacteria. This suggests that the history of humans consuming nisin is older than the tradition of consuming fermented milk products.

  16. Physiological Studies of Lactococcus lactis

    DEFF Research Database (Denmark)

    Hansen, Gunda

    fermentation, downstream processing and storage in the absence of a protectant. However, storing freeze-dried L. lactis cells at 30 °C negatively affected the culturability and acidification activity. The reactivation of freeze-dried cells in fermentation medium prior to flow cytometric viability assessment...

  17. Increased production of folate by metabolic engineering of Lactococcus lactis

    NARCIS (Netherlands)

    Sybesma, W.F.H.; Starrenburg, M.; Kleerebezem, M.; Mierau, I.; Vos, de W.M.; Hugenholtz, J.

    2003-01-01

    The dairy starter bacterium Lactococcus lactis is able to synthesize folate and accumulates large amounts of folate, predominantly in the polyglutamyl form. Only small amounts of the produced folate are released in the extracellular medium. Five genes involved in folate biosynthesis were identified

  18. Structure-function analysis of multidrug transporters in Lactococcus lactis

    NARCIS (Netherlands)

    van Veen, HW; Putman, M; Margolles, A; Sakamoto, K; Konings, WN

    1999-01-01

    The active extrusion of cytotoxic compounds from the cell by multidrug transporters is one of the major causes of failure of chemotherapeutic treatment of tumor cells and of infections by pathogenic microorganisms. A multidrug transporter in Lactococcus lactis, LmrA, is a member of the ATP-binding c

  19. Lactococcus lactis as host for overproduction of functional membrane proteins

    NARCIS (Netherlands)

    Kunji, ERS; Slotboom, DJ; Poolman, B

    2003-01-01

    Lactococcus lactis has many properties that are ideal for enhanced expression of membrane proteins. The organism is easy and inexpensive to culture, has a single membrane and relatively mild proteolytic activity. Methods for genetic manipulation are fully established and a tightly controlled promote

  20. Multidrug transporters and antibiotic resistance in Lactococcus lactis

    NARCIS (Netherlands)

    Poelarends, GJ; Mazurkiewicz, P; Konings, WN

    2002-01-01

    The Gram-positive bacterium Lactococcus lactis produces two distinct multidrug transporters, designated LmrA and LmrP, that both confer resistance to a wide variety of cationic lipophilic cytotoxic compounds as well as to many clinically relevant antibiotics. While LmrP is a proton/drug antiporter t

  1. The effect of co-administration of lactobacillus probiotics and bifidobacterium on spatial memory and learning in diabetic rats

    Directory of Open Access Journals (Sweden)

    H Alaei

    2012-12-01

    Full Text Available Background: Diabetes mellitus affects numerous intracellular metabolic processes, which are reflected by changes in the concentration of some plasma constituents. Particularly, the disease may indirectly undermine some functions of the nervous system including learning and memory through altering oxidative stress status. On the other hand, probiotics can enhance the antioxidant capacity. This study was designed to evaluate the effects of probiotics on spatial memory, maze learning and indices of oxidative stress in diabetic rats.Methods: In this experimental study, 40 male Wistar rats were randomly allocated to 4 groups (n=10 for each: Control (CO, Control probiotic (CP, Control diabetic (DC, and Diabetic probiotic (DP. The probiotic supplement, including Lactobacillus acidophilus, Lactobacillus fermentum, Bifidobacterium lactis (334 mg of each with a CFU of ~1010, was administered through drinking water every 12 hours for 8 weeks. Using morris water maze (MWM, spatial learning and memory were evaluated. Serum insulin and oxidative stress indices, including superoxide dismutase (SOD and 8-hydroxy-2'-deoxyguanosine (8-OHdG, were measured by standard laboratory kits.Results: Oral administration of probiotics improved impairment of spatial learning (P=0.008 and consolidated memory (P=0.01 in the rats. Moreover, probiotic treatment increased serum insulin (P<0.0001 and serum superoxide dismutase activity (P=0.007 while it decreased their blood glucose (P=0.006 and 8-OHdG (P<0.0001.Conclusion: Probiotic supplementation reversed the serum concentrations of insulin and glucose along with an increase in antioxidant capacity in diabetic rats. It also improved spatial learning and memory in the animals. Relevancy of the metabolic changes and behavioral functions need to be further studied.

  2. Purification and Characterization of Conjugated Bile Salt Hydrolase from Bifidobacterium longum BB536

    OpenAIRE

    Grill, J; Schneider, F.; Crociani, J.; Ballongue, J.

    1995-01-01

    Bifidobacterium species deconjugate taurocholic, taurodeoxycholic, taurochenodeoxycholic, glycocholic, glycodeoxycholic, and glycochenodeoxycholic acids. The enzyme level increases in the growth phase. No increase in activity is observed for the cytoplasmic enzyme after addition of conjugated bile acids to a stationary-phase culture. Conjugated bile salt hydrolase (BSH) was purified from Bifidobacterium longum BB536. Its apparent molecular mass in denaturing polyacrylamide gel electrophoresis...

  3. A Bifidobacterium mixed-species microarray for high resolution discrimination between intestinal bifidobacteria

    NARCIS (Netherlands)

    Boesten, R.J.; Schuren, F.H.; Vos, de W.M.

    2009-01-01

    A genomic DNA-based microarray was constructed containing over 6000 randomly cloned genomic fragments of approximately 1-2 kb from six mammalian intestinal Bifidobacterium spp. including B. adolescentis, B. animalis, B. bifidum, B. catenulatum, B. longum and B. pseudolongum. This Bifidobacterium Mix

  4. A convenient and reproducible method to genetically transform bacteria of the genus Bifidobacterium

    NARCIS (Netherlands)

    Argnani, A.; Leer, R.J.; Luijk, N. van; Pouwels, P.H.

    1996-01-01

    A protocol was developed for the introduction of foreign plasmid DNA into various Bifidobacterium strains. The method, which is applicable to all Bifidobacterium species tested so far, is based on electroporation of bacteria made competent by preincubation in electroporation buffer for several hours

  5. Lactose-mediated carbon catabolite repression of putrescine production in dairy Lactococcus lactis is strain dependent.

    Science.gov (United States)

    del Rio, Beatriz; Ladero, Victor; Redruello, Begoña; Linares, Daniel M; Fernández, Maria; Martín, Maria Cruz; Alvarez, Miguel A

    2015-06-01

    Lactococcus lactis is the lactic acid bacterial (LAB) species most widely used as a primary starter in the dairy industry. However, several strains of L. lactis produce the biogenic amine putrescine via the agmatine deiminase (AGDI) pathway. We previously reported the putrescine biosynthesis pathway in L. lactis subsp. cremoris GE2-14 to be regulated by carbon catabolic repression (CCR) via glucose but not lactose (Linares et al., 2013). The present study shows that both these sugars repress putrescine biosynthesis in L. lactis subsp. lactis T3/33, a strain isolated from a Spanish artisanal cheese. Furthermore, we demonstrated that both glucose and lactose repressed the transcriptional activity of the aguBDAC catabolic genes of the AGDI route. Finally, a screening performed in putrescine-producing dairy L. lactis strains determined that putrescine biosynthesis was repressed by lactose in all the L. lactis subsp. lactis strains tested, but in only one L. lactis subsp. cremoris strain. Given the obvious importance of the lactose-repression in cheese putrescine accumulation, it is advisable to consider the diversity of L. lactis in this sense and characterize consequently the starter cultures to select the safest strains.

  6. Modeling Lactococcus lactis using a genome-scale flux model

    Directory of Open Access Journals (Sweden)

    Nielsen Jens

    2005-06-01

    Full Text Available Abstract Background Genome-scale flux models are useful tools to represent and analyze microbial metabolism. In this work we reconstructed the metabolic network of the lactic acid bacteria Lactococcus lactis and developed a genome-scale flux model able to simulate and analyze network capabilities and whole-cell function under aerobic and anaerobic continuous cultures. Flux balance analysis (FBA and minimization of metabolic adjustment (MOMA were used as modeling frameworks. Results The metabolic network was reconstructed using the annotated genome sequence from L. lactis ssp. lactis IL1403 together with physiological and biochemical information. The established network comprised a total of 621 reactions and 509 metabolites, representing the overall metabolism of L. lactis. Experimental data reported in the literature was used to fit the model to phenotypic observations. Regulatory constraints had to be included to simulate certain metabolic features, such as the shift from homo to heterolactic fermentation. A minimal medium for in silico growth was identified, indicating the requirement of four amino acids in addition to a sugar. Remarkably, de novo biosynthesis of four other amino acids was observed even when all amino acids were supplied, which is in good agreement with experimental observations. Additionally, enhanced metabolic engineering strategies for improved diacetyl producing strains were designed. Conclusion The L. lactis metabolic network can now be used for a better understanding of lactococcal metabolic capabilities and potential, for the design of enhanced metabolic engineering strategies and for integration with other types of 'omic' data, to assist in finding new information on cellular organization and function.

  7. Use of a genetically enhanced, pediocin-producing starter culture, Lactococcus lactis subsp. lactis MM217, to control Listeria monocytogenes in cheddar cheese

    NARCIS (Netherlands)

    Buyong, N; Kok, J; Luchansky, JB

    1998-01-01

    Cheddar cheese was prepared with Lactococcus lactis subsp, lactis MM217, a starter culture which contains pMC117 coding for pediocin PA-1, About 75 liters of pasteurized milk (containing ca, 3.6% fat) was inoculated with strain MM217 (ca, 10(6) CFU per ml) and a mixture of three Listeria monocytogen

  8. Effect of inulin on the human gut microbiota: stimulation of Bifidobacterium adolescentis and Faecalibacterium prausnitzii.

    Science.gov (United States)

    Ramirez-Farias, Carlett; Slezak, Kathleen; Fuller, Zoë; Duncan, Alan; Holtrop, Grietje; Louis, Petra

    2009-02-01

    Prebiotics are food ingredients that improve health by modulating the colonic microbiota. The bifidogenic effect of the prebiotic inulin is well established; however, it remains unclear which species of Bifidobacterium are stimulated in vivo and whether bacterial groups other than lactic acid bacteria are affected by inulin consumption. Changes in the faecal microbiota composition were examined by real-time PCR in twelve human volunteers after ingestion of inulin (10 g/d) for a 16-d period in comparison with a control period without any supplement intake. The prevalence of most bacterial groups examined did not change after inulin intake, although the low G+C % Gram-positive species Faecalibacterium prausnitzii exhibited a significant increase (10.3% for control period v. 14.5% during inulin intake, P=0.019). The composition of the genus Bifidobacterium was studied in four of the volunteers by clone library analysis. Between three and five Bifidobacterium spp. were found in each volunteer. Bifidobacterium adolescentis and Bifidobacterium longum were present in all volunteers, and Bifidobacterium pseudocatenulatum, Bifidobacterium animalis, Bifidobacterium bifidum and Bifidobacterium dentium were also detected. Real-time PCR was employed to quantify the four most prevalent Bifidobacterium spp., B. adolescentis, B. longum, B. pseudocatenulatum and B. bifidum, in ten volunteers carrying detectable levels of bifidobacteria. B. adolescentis showed the strongest response to inulin consumption, increasing from 0.89 to 3.9% of the total microbiota (P=0.001). B. bifidum was increased from 0.22 to 0.63% (P<0.001) for the five volunteers for whom this species was present.

  9. Expression of Helicobacter pylori hspA Gene in Lactococcus lactis NICE System and Experimental Study on Its Immunoreactivity

    OpenAIRE

    Xiao-Juan Zhang; Shu-Ying Feng; Zhi-Tao Li; Yan-Ming Feng

    2015-01-01

    Aim. The aim of this study was to develop an oral Lactococcus lactis (L. lactis) vaccine against Helicobacter pylori (H. pylori). Methods. After L. lactis NZ3900/pNZ8110-hspA was constructed, growth curves were plotted to study whether the growth of recombinant L. lactis was affected after hspA was cloned into L. lactis and whether the growth of empty bacteria, empty plasmid bacteria, and recombinant L. lactis was affected by different concentrations of Nisin; SDS-PAGE and Western blot were a...

  10. Cell wall anchoring of the Campylobacter antigens to Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    Patrycja Anna Kobierecka

    2016-02-01

    Full Text Available Campylobacter jejuni is the most frequent cause of human food-borne gastroenteritis and chicken meat is the main source of infection. Recent studies showed that broiler chicken immunization against Campylobacter should be the most efficient way to lower the number of human infections by this pathogen. Induction of the mucosal immune system after oral antigen administration should provide protective immunity to chickens. In this work we tested the usefulness of Lactococcus lactis, the most extensively studied lactic acid bacterium, as a delivery vector for Campylobacter antigens. First we constructed hybrid protein – CjaA antigen presenting CjaD peptide epitopes on its surface. We showed that specific rabbit anti-rCjaAD serum reacted strongly with both CjaA and CjaD produced by a wild type Campylobacter jejuni strain. Next, rCjaAD and CjaA were fused to the C-terminus of the L. lactis YndF containing the LPTXG motif. The genes expressing these proteins were transcribed under control of the L. lactis Usp45 promoter and their products contain the Usp45 signal sequences. This strategy ensures a cell surface location of both analysed proteins, which was confirmed by immunofluorescence assay. In order to evaluate the impact of antigen location on vaccine prototype efficacy, a L. lactis strain producing cytoplasm-located rCjaAD was also generated. Animal experiments showed a decrease of Campylobacter cecal load in vaccinated birds as compared with the control group and showed that the L. lactis harboring the surface-exposed rCjaAD antigen afforded greater protection than the L. lactis producing cytoplasm-located rCjaAD. To the best of our knowledge, this is the first attempt to employ LAB (Lactic Acid Bacteria strains as a mucosal delivery vehicle for chicken immunization. Although the observed reduction of chicken colonization by Campylobacter resulting from vaccination was rather moderate, the experiments showed that LAB strains can be considered

  11. Generation of Dipeptidyl Peptidase-IV-Inhibiting Peptides from β-Lactoglobulin Secreted by Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    Suguru Shigemori

    2014-01-01

    Full Text Available Previous studies showed that hydrolysates of β-lactoglobulin (BLG prepared using gastrointestinal proteases strongly inhibit dipeptidyl peptidase-IV (DPP-IV activity in vitro. In this study, we developed a BLG-secreting Lactococcus lactis strain as a delivery vehicle and in situ expression system. Interestingly, trypsin-digested recombinant BLG from L. lactis inhibited DPP-IV activity, suggesting that BLG-secreting L. lactis may be useful in the treatment of type 2 diabetes mellitus.

  12. Lactococcus lactis-based vaccines from laboratory bench to human use: an overview.

    Science.gov (United States)

    Bahey-El-Din, Mohammed

    2012-01-17

    Developing effective vaccines is an important weapon in the battle against potential pathogens and their evolving antibiotic resistance trends. Several vaccine delivery vectors have been investigated among which the generally regarded as safe (GRAS) Lactococcus lactis has a distinguished position. In this review, different factors affecting the efficacy of L. lactis-based vaccines are discussed. In addition, the issues of biological containment and pharmaceutical quality assurance of L. lactis vaccines are highlighted. These issues are critical for the success of medical translation of L. lactis-based vaccines from research laboratories to clinical use by ensuring consistent manufacturing of safe and efficacious vaccines.

  13. 青春双歧杆菌对2型糖尿病模型大鼠肠道菌群和脂质代谢的影响%Influence of Bifidobacterium adolescentis on intestinal flora and lipid metabolism in type 2 diabetes mellitus rats

    Institute of Scientific and Technical Information of China (English)

    刘伯阳; 姚淑娟; 夏美玲

    2009-01-01

    目的 通过观察青春双歧杆菌对2型糖尿病模型大鼠肠道菌群的变化,和血清中总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL-C)、超氧化物歧化酶(SOD)和丙二醛(MDA)的水平,探讨青春双歧杆菌对2型糖尿病模型大鼠肠道功能和脂质代谢的影响.方法 采用青春双歧杆菌灌胃2型糖尿病模型大鼠,取粪便检查正常菌群,取血和脏器检测TC、TG、HDL-C、SOD和MDA含量.结果 青春双歧杆菌导致肠道内双歧杆菌、乳杆菌的数量增加,而肠杆菌、肠球菌数量下降;TC、TG和MDA水平下降,而HDL-C和SOD水平升高.结论 青春双歧杆菌具有改善2型糖尿病模型大鼠肠道功能和降血脂作用,与二甲双胍联合应用效果更佳.%Objective To observe the change of intestinal flora caused by Bifidobacterium adolescentis and the levels of total cholesterol, Triglyceride, high density lipoprotein, superoxide dismutase and malondialdehyde in type 2 diabetes mellitus rats,and explore the influence of Bifidobacterium adolescentis on intestinal flora and lipid metabolism in type 2 diabetes mellitus rats. Method The levels of normal flora, total cholesterol, Triglyceride, high density lipoprotein,superoxide dismutase and malondialdehyde were examined after gavaging type 2 diabetes mellitus rats by Bifidobacterium adolescentis. Result Bifidobacterium adolescentis had caused the number of bifidobacterium and lactis to increase while that of Enter-obacter and Enterococcus to decrease;The levels of total cholesterol, Triglyceride and malondialdehyde were reduced while those of high density lipoprotein and superoxide dismutase were improved. Conclusion Bifidobacterium adolescentis has the effect of improving intestinal function and lowering serum lipids in type 2 diabetes mellitus rats,the effect of which can be improved when combined with metformin.

  14. Characterization of fructose 6 phosphate phosphoketolases purified from Bifidobacterium species.

    Science.gov (United States)

    Grill, J P; Crociani, J; Ballongue, J

    1995-07-01

    Fructose 6 phosphate phosphoketolases (F6PPKs) were purified from Bifidobacterium longum BB536, B. dentium ATCC 27534, B. globosum ATCC 25864, and Bifidobacterium animalis ATCC 25527. Concerning ions (Cu++, Zn++, Ca++, Mg++, Fe++, Co++, Mn++) and common enzyme inhibitors (fructose, ammonium sulfate, iodoacetate, and parachloromercuribenzoic acid), no difference appeared between the enzymes. Cu++, parachloromercuribenzoic acid (pCMB), and mercuric acetate induced high enzymatic inhibition. The study of pCMB demonstrated a noncompetitive inhibition. Additional results showed that the sulfhydryl group was not involved in catalytic reaction. Photooxidation experiments and determination of ionizable group pKas (5.16-7.17) suggested the presence of one or more histidines necessary for the catalytic reaction and explained the inhibition observed with pCMB. In light of the noncompetitive inhibition, this group was not directly involved in substrate binding. Determination of Km demonstrated that the affinities for fructose 6 phosphate in the case of animal and human origin strains were close. In addition, the same enzymatic efficiency (Kcat/Km) was obtained for each strain. The F6PPK activity was regulated by sodium pyrophosphate, ATP, and especially by ADP.

  15. Bioconversion of soy isoflavones daidzin and daidzein by Bifidobacterium strains.

    Science.gov (United States)

    Raimondi, Stefano; Roncaglia, Lucia; De Lucia, Marzia; Amaretti, Alberto; Leonardi, Alan; Pagnoni, Ugo Maria; Rossi, Maddalena

    2009-01-01

    Twenty-two strains of Bifidobacterium, representative of eight major species of human origin, were screened for their ability to transform the isoflavones daidzin and daidzein. Most of the strains released the aglycone from daidzin and 12 gave yields higher than 90%. The kinetics of growth, daidzin consumption, and daidzein production indicated that the hydrolytic activity occurred during the growth. The supernatant of the majority of the strains did not release the aglycone from daidzin, suggesting that cell-associated beta-glucosidases (beta-Glu) are mainly responsible for the metabolism of soybean glyco-conjugates. Cell-associated beta-Glu was mainly intracellular and significantly varied among the species and the strains. The lack of beta-Glu was correlated with the inability to hydrolyze daidzin. Although S-equol production by anaerobic intestinal bacteria has been established, information on S-equol-producing bifidobacteria is contradictory. In this study, 22 bifidobacteria failed to transform daidzein into reduced metabolites under all the experimental conditions, excluding any role in the reductive pathway of daidzein toward the production of S-equol. These results suggest that selected probiotic strains of Bifidobacterium can be used to speed up the release of daidzein, improving its bioavailability for absorption by colonic mucosa and/or biotransformation to S-equol by other intestinal microorganisms.

  16. Experimental determination of control of glycolysis in Lactococcus lactis

    DEFF Research Database (Denmark)

    Købmann, Brian Jensen; Andersen, Heidi Winterberg; Solem, Christian

    2002-01-01

    and below the wild-type level. We review a genetic approach that is well suited for both Metabolic Optimization and Metabolic Control Analysis and studies on the importance of a number of glycolytic enzymes for metabolic fluxes in Lactococcus lactis. The glycolytic enzymes phosphofructokinase (PFK......The understanding of control of metabolic processes requires quantitative studies of the importance of the different enzymatic steps for the magnitude of metabolic fluxes and metabolite concentrations. An important element in such studies is the modulation of enzyme activities in small steps above......), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), pyruvate kinase (PYK) and lactate dehydrogenase (LDH) are shown to have no significant control on the glycolytic flux in exponentially growing cells of L. lactis MG1363. Introduction of an uncoupled ATPase activity results in uncoupling of glycolysis from biomass...

  17. Plasma mutation breeding of Lactococcus lactis in producing Nisin

    Directory of Open Access Journals (Sweden)

    Shuanli ZHANG

    2015-12-01

    Full Text Available With Nisin-producing Lactococcus lactis as the starting strain, the strain with tolerance to 10 000 IU/mL Nisin is selected on high-concentration Nisin medium. The Nisin titer of the strain is up to 1 680 IU/mL. As the starting strain, the strain is further treated by atmospheric and room temperature plasmas(ARTP and mutant strain for high yield of Nisin is quickly selected with 24 well culture plate. At a survival rate of 3%, the positive mutation rate of the Lactococcus lactis is 273% compared with the starting strain. The results of shake flask culture further confirmed that one positive mutant strains could produce 6 120 IU/mL Nisin.

  18. Proteome analysis of the purine stimulon from Lactococcus lactis

    DEFF Research Database (Denmark)

    Beyer, N.H.; Roepstorff, P.; Hammer, Karin;

    2003-01-01

    Box-Pribnov box structures resembling the PurR activated promoters for the purDEK and purCSQLF operons. Most, and possibly all members of the Psu stimulon are thus members of the PurR regulon. Five Psu proteins could not be identified. The second stimulon, the Psd stimulon (purine starvation decreased), whose...... escaped identification. The last, Dcu (decoynine up-regulated), stimulon contained proteins whose synthesis escaped the severe general depression during inhibition of the GMP synthetase by decoynine. This regulon was comprised of mostly glycolytic enzymes (fructose bisphosphate aldolase, enolase, pyruvate...... kinase) and translation elongation factors (GTPases: EF-TU, EF-G). Two Dcu proteins could not be identified. Out of 28 proteins subjected to mass spectrometry, 19 could be readily identified despite the fact that only the genome sequence of a strain of L. lactis subsp. lactis was available. The two...

  19. Genes but not genomes reveal bacterial domestication of Lactococcus lactis.

    Directory of Open Access Journals (Sweden)

    Delphine Passerini

    Full Text Available BACKGROUND: The population structure and diversity of Lactococcus lactis subsp. lactis, a major industrial bacterium involved in milk fermentation, was determined at both gene and genome level. Seventy-six lactococcal isolates of various origins were studied by different genotyping methods and thirty-six strains displaying unique macrorestriction fingerprints were analyzed by a new multilocus sequence typing (MLST scheme. This gene-based analysis was compared to genomic characteristics determined by pulsed-field gel electrophoresis (PFGE. METHODOLOGY/PRINCIPAL FINDINGS: The MLST analysis revealed that L. lactis subsp. lactis is essentially clonal with infrequent intra- and intergenic recombination; also, despite its taxonomical classification as a subspecies, it displays a genetic diversity as substantial as that within several other bacterial species. Genome-based analysis revealed a genome size variability of 20%, a value typical of bacteria inhabiting different ecological niches, and that suggests a large pan-genome for this subspecies. However, the genomic characteristics (macrorestriction pattern, genome or chromosome size, plasmid content did not correlate to the MLST-based phylogeny, with strains from the same sequence type (ST differing by up to 230 kb in genome size. CONCLUSION/SIGNIFICANCE: The gene-based phylogeny was not fully consistent with the traditional classification into dairy and non-dairy strains but supported a new classification based on ecological separation between "environmental" strains, the main contributors to the genetic diversity within the subspecies, and "domesticated" strains, subject to recent genetic bottlenecks. Comparison between gene- and genome-based analyses revealed little relationship between core and dispensable genome phylogenies, indicating that clonal diversification and phenotypic variability of the "domesticated" strains essentially arose through substantial genomic flux within the dispensable

  20. Mobile CRISPR/Cas-mediated bacteriophage resistance in Lactococcus lactis.

    Directory of Open Access Journals (Sweden)

    Anne M Millen

    Full Text Available Lactococcus lactis is a biotechnological workhorse for food fermentations and potentially therapeutic products and is therefore widely consumed by humans. It is predominantly used as a starter microbe for fermented dairy products, and specialized strains have adapted from a plant environment through reductive evolution and horizontal gene transfer as evidenced by the association of adventitious traits with mobile elements. Specifically, L. lactis has armed itself with a myriad of plasmid-encoded bacteriophage defensive systems to protect against viral predation. This known arsenal had not included CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins, which forms a remarkable microbial immunity system against invading DNA. Although CRISPR/Cas systems are common in the genomes of closely related lactic acid bacteria (LAB, none was identified within the eight published lactococcal genomes. Furthermore, a PCR-based search of the common LAB CRISPR/Cas systems (Types I and II in 383 industrial L. lactis strains proved unsuccessful. Here we describe a novel, Type III, self-transmissible, plasmid-encoded, phage-interfering CRISPR/Cas discovered in L. lactis. The native CRISPR spacers confer resistance based on sequence identity to corresponding lactococcal phage. The interference is directed at phages problematic to the dairy industry, indicative of a responsive system. Moreover, targeting could be modified by engineering the spacer content. The 62.8-kb plasmid was shown to be conjugally transferrable to various strains. Its mobility should facilitate dissemination within microbial communities and provide a readily applicable system to naturally introduce CRISPR/Cas to industrially relevant strains for enhanced phage resistance and prevention against acquisition of undesirable genes.

  1. Therapeutic drug delivery by genetically modified Lactococcus lactis.

    Science.gov (United States)

    Steidler, Lothar; Rottiers, Pieter

    2006-08-01

    Food-grade bacteria have been consumed throughout history without associated pathologies and are, therefore, absolutely safe to ingest. Unexpectedly, Lactococcus lactis (L. lactis), known from cheese production, can be genetically engineered to constantly secrete satisfactory amounts of bioactive cytokines. Both of these features enabled the development of a new kind of topical delivery system: topical and active delivery of therapeutic proteins by genetically modified micro-organisms. The host organism's record inspired the development of applications that target intestinal diseases. In a variety of mouse models, chronic colon inflammation can be successfully treated with (interleukin) IL-10-secreting L. lactis. Trefoil factor (TFF) producer strains have also been shown to be very effective in the treatment of acute colitis. Such novel therapeutic strains are textbook examples of genetically modified (GM) organisms. There are legitimate concerns with regard to the deliberate release of GM micro-organisms. On development of these applications, therefore, we have engineered these bacteria in such a way that biological containment is guaranteed. The essential gene thyA, encoding thymidylate synthase, has been exchanged for IL-10. This makes the GM strain critically dependent on thymidine. Lack of thymidine, for example, resulting from thymidine consumption by thyA-deficient strains-will irreversibly lead to induced "thymidine-less death." This accomplishment has created the possibility of using this strategy for application in human medicine.

  2. Recombinant expression of Laceyella sacchari thermitase in Lactococcus lactis.

    Science.gov (United States)

    Jørgensen, Casper M; Madsen, Søren M; Vrang, Astrid; Hansen, Ole C; Johnsen, Mads G

    2013-12-01

    Thermitase (EC 3.4.21.66) is a thermostable endo-protease with the ability to convert various food relevant substrates into low-molecular weight peptides. A thermitase produced by Laceyella sacchari strain DSM43353 was found to have a mature amino acid sequence nearly identical to that of the original thermitase isolated from Thermoactinomyces vulgaris. The DSM43353 thermitase gene sequence contains a pro-peptide including parts of an I9 inhibitor motif. Expression of the thermitase gene in the Lactococcus lactis P170 expression system allowed secretion of stable thermitase in an auto-induced fermentation setup at 30°C. Thermitase accumulated in the culture supernatant during batch fermentations and was easily activated at 50°C or by prolonged dialysis. The activation step resulted in an almost complete degradation of endogenous L. lactis host proteins present in the supernatant. Mature activated product was stable at 50°C and functional at pH values between pH 6 and pH 11, suggesting that substrate hydrolysis can be performed over a broad range of pH values. The L. lactis based P170 expression system is a simple and safe system for obtaining food compatible thermitase in the range of 100 mg/L.

  3. Emulsifying, rheological and physicochemical properties of exopolysaccharide produced by Bifidobacterium longum subsp. infantis CCUG 52486 and Bifidobacterium infantis NCIMB 702205.

    Science.gov (United States)

    Prasanna, P H P; Bell, A; Grandison, A S; Charalampopoulos, D

    2012-09-01

    The rheological, emulsification and certain physicochemical properties of purified exopolysaccharides (EPS) of Bifidobacterium longum subsp. infantis CCUG 52486 and Bifidobacterium infantis NCIMB 702205 were studied and compared with those of guar gum and xanthan gum. The two strains were grown in skim milk supplemented with 1.5% (w/v) casein hydrolysate at 37 °C for 24h; they both produced heteropolysaccharides with different molecular mass and composition. The carbohydrate content of both polymers was more than 92% and no protein was detected. The EPS of B. longum subsp. infantis CCUG 52486 showed highly branched entangled porous structure under scanning electron microscopy. Higher intrinsic viscosity was observed for the EPS of B. longum subsp. infantis CCUG 52486 compared to the EPS of B. infantis NCIMB 702205 and guar gum. Both polymers showed pseudoplastic non-Newtonian fluid behaviour in an aqueous solution. The EPS of B. infantis NCIMB 702205 and B. longum subsp. infantis CCUG 52486 produced more stable emulsions with orange oil, sunflower seed oil, coconut oil and xylene compared to guar and xanthan gum. The EPS of B. longum subsp. infantis CCUG 52486 is the most promising one for applications in the food industry, as it had higher intrinsic viscosity, higher apparent viscosity in aqueous solution, porous dense entangled structure and good emulsification activity.

  4. Identification and characterization of the alpha-acetolactate synthase gene from Lactococcus lactis subsp. lactis biovar diacetylactis.

    Science.gov (United States)

    Marugg, J D; Goelling, D; Stahl, U; Ledeboer, A M; Toonen, M Y; Verhue, W M; Verrips, C T

    1994-01-01

    The conversion of 3-13C-labelled pyruvate in an acetoin-producing clone from a Lactococcus lactis subsp. lactis biovar diacetylactis strain DSM 20384 plasmid bank in Escherichia coli was studied by 13C nuclear magnetic resonance analysis. The results showed that alpha-acetolactate was the first metabolic product formed from pyruvate, whereas acetoin appeared at a much slower rate and reached only low concentrations. This alpha-acetolactate production shows that the cells express the gene for alpha-acetolactate synthase (als). Nucleotide sequence analysis identified an open reading frame encoding a protein of 554 amino acids. The deduced amino acid sequence exhibits extensive similarities to those of known alpha-acetolactate synthases from both prokaryotes and eukaryotes. The als gene is expressed on a monocistronic transcriptional unit, which is transcribed from a promoter located just upstream of the coding region. Images PMID:8017926

  5. Recombinant invasive Lactococcus lactis can transfer DNA vaccines either directly to dendritic cells or across an epithelial cell monolayer

    NARCIS (Netherlands)

    Azevedo, de Marcela; Meijerink, Marjolein; Taverne, Nico; Pereira, Vanessa Bastos; LeBlanc, Jean Guy; Azevedo, Vasco; Miyoshi, Anderson; Langella, Philippe; Wells, J.M.; Chatel, Jean Marc

    2015-01-01

    Lactococcus lactis (L. lactis), a generally regarded as safe (GRAS) bacterium has recently been investigated as a mucosal delivery vehicle for DNA vaccines. Because of its GRAS status, L. lactis represents an attractive alternative to attenuated pathogens. Previous studies showed that eukaryotic

  6. EXPRESSION OF A CHITINASE GENE FROM SERRATIA-MARCESCENS IN LACTOCOCCUS-LACTIS AND LACTOBACILLUS-PLANTARUM

    NARCIS (Netherlands)

    BRURBERG, MB; HAANDRIKMAN, AJ; LEENHOUTS, KJ; VENEMA, G; NES, IF

    1994-01-01

    A chitinase gene from the Gram-negative bacterium Serratia marcescens BJL200 was cloned in Lactococcus lactis subsp. lactis MG1363 and in the silage inoculum strain Lactobacillus plantarum E19b. The chitinase gene was expressed as an active enzyme at a low level in Lactococcus lactis, when cloned in

  7. Hydrophobic membrane thickness and lipid-protein interactions of the leucine transport system of Lactococcus lactis

    NARCIS (Netherlands)

    in t Veld, Gerda; Driessen, Arnold J.M.; Kamp, Jos A.F. op den; Konings, Wil N.

    1991-01-01

    The effect of the phospholipid acyl chain carbon number on the activity of the branched-chain amino acid transport system of Lactococcus lactis has been investigated. Major fatty acids identified in a total lipid extract of L. lactis membranes are palmitic acid (16:0), oleic acid (18:1) and the cycl

  8. Complete genome sequence of Lactococcus lactis S0, an efficient producer of nisin.

    Science.gov (United States)

    Zhao, Fangyuan; Ma, Hongchu; Lu, Ying; Teng, Kunling; Kang, Xusheng; Wang, Fangfang; Yang, Xiaopan; Zhong, Jin

    2015-03-20

    Lactococcus lactis S0 is a nisin Z-producing strain isolated from milk, and the nisin production of the strain can reach 4000 IU/ml under fermenting condition. Here, we present the complete genome sequence of L. lactis S0 which includes a single circular chromosome.

  9. Morphology, genome sequence, and structural proteome of type phage P335 from Lactococcus lactis

    DEFF Research Database (Denmark)

    Labrie, Simon J.; Josephsen, Jytte; Neve, Horst;

    2008-01-01

    Lactococcus lactis phage P335 is a virulent type phage for the species that bears its name and belongs phage P335 is a virulent type phage for the species that bears its name and belongs to the Siphoviridae family. Morphologically, P335 resembled the L. lactis phages TP901-1 and Tuc2009, except...

  10. Characteristics and Osmoregulatory Roles of Uptake Systems for Proline and Glycine Betaine in Lactococcus lactis

    NARCIS (Netherlands)

    Molenaar, Douwe; Hagting, Anja; Alkema, Harmen; Driessen, Arnold J.M.; Konings, Wilhelmus

    1993-01-01

    Lactococcus lactis subsp. lacti ML3 contains high pools of proline or betaine when grown under conditions of high osmotic strength. These pools are created by specific transport systems. A high-affinity uptake system for glycine betaine (betaine) with a Km of 1.5 µM is expressed constitutively. The

  11. Time-resolved genetic responses of Lactococcus lactis to a dairy environment

    NARCIS (Netherlands)

    Bachmann, H.; Wilt, de L.; Kleerebezem, M.; Hylckama Vlieg, van J.E.T.

    2010-01-01

    Lactococcus lactis is one of main bacterial species found in mixed dairy starter cultures for the production of semi-hard cheese. Despite the appreciation that mixed cultures are essential for the eventual properties of the manufactured cheese the vast majority of studies on L. lactis were carried o

  12. Heterologous expression and characterization of recombinant Lactococcus lactis neutral endopeptidase (Neprilysin)

    NARCIS (Netherlands)

    Lian, W; Wu, D; Konings, W.N; Mierau, I; Hersh, L.B

    1996-01-01

    A neutral endopeptidase (NEP) from Lactococcus lactis has recently been cloned and shown to contain high sequence homology with the human neutral endopeptidase, endopeptidase 24.11 (I. Mierau et al., J. Bacteriol. 175, 2087-2096, 1993). The gene for the neutral endopeptidase from L. lactis was clone

  13. Molecular analysis of the replication origin of the Lactococcus lactis plasmid pCI305

    NARCIS (Netherlands)

    Foley, S; Bron, S; Venema, G; Daly, C; Fitzgerald, GF

    1996-01-01

    The replication origin region, ori, of the Lactococcus lactis subsp. lactis plasmid pCI305 contains three-and-one-half directly repeated 22-bp sequences and two inverted repeat sequences, IR1 and IR2. These inverted repeat sequences overlap the promoter of the repB gene, which encodes a protein (Rep

  14. Arginine transport in Streptococcus lactis is catalyzed by a cationic exchanger

    NARCIS (Netherlands)

    Driessen, Arnold J.M.; Poolman, Bert; Kiewiet, Rense; Konings, Wil N.

    1987-01-01

    Streptococcus lactis metabolizes arginine via the arginine deiminase pathway to ornithine, CO2, NH3, and ATP. The translocation of arginine and ornithine has been studied using membrane vesicles of galactose/arginine-grown cells of S. lactis fused with cytochrome c oxidase proteoliposomes by the fre

  15. Transcriptomic profile of aguR deletion mutant of Lactococcus lactis subsp. cremoris CECT 8666

    NARCIS (Netherlands)

    Del Rio, Beatriz; Linares, Daniel M; Redruello, Begoña; Martin, Maria Cruz; Fernandez, Maria; de Jong, Anne; Kuipers, Oscar P; Ladero, Victor; Alvarez, Miguel A

    2015-01-01

    Lactococcus lactis subsp. cremoris CECT 8666 (formerly GE2-14) is a dairy strain that catabolizes agmatine (a decarboxylated derivative of arginine) into the biogenic amine putrescine by the agmatine deiminase (AGDI) pathway [1]. The AGDI cluster of L. lactis is composed by five genes aguR, aguB, ag

  16. Regulation of Arginine-Ornithine Exchange and the Arginine Deiminase Pathway in Streptococcus lactis

    NARCIS (Netherlands)

    POOLMAN, B; DRIESSEN, AJM; KONINGS, WN

    1987-01-01

    Streptococcus lactis metabolizes arginine by the argiqine deiminase (ADI) pathway. Resting cells of S. lactis grown in the presence of galactose and arginine maintain a high intracellular ornithine pool in the absence of arginine and other exogenous energy sources. Addition of arginine results in a

  17. Genotype-phenotype matching analysis of 38 Lactococcus lactis strains using random forest methods

    NARCIS (Netherlands)

    Bayjanov, J.; Starrenburg, M.J.; Sijde, M.R. van der; Siezen, R.J.; Hijum, S.A.F.T. van

    2013-01-01

    BACKGROUND: Lactococcus lactis is used in dairy food fermentation and for the efficient production of industrially relevant enzymes. The genome content and different phenotypes have been determined for multiple L. lactis strains in order to understand intra-species genotype and phenotype diversity a

  18. Genetically modified lactococcus lactis for delivery of human interleukin-10 to dendritic cells

    NARCIS (Netherlands)

    H. Braat (Henri); I.L. Huibregtse (Inge ); S.A.J. Zaat (Sebastiaan); M.L. Kapsenberg (Martien ); M.A. Sartori da Silva; M.P. Peppelenbosch (Maikel); S. van Deventer (Sander)

    2012-01-01

    textabstractInterleukin-10 (IL-10) plays an indispensable role in mucosal tolerance by programming dendritic cells (DCs) to induce suppressor Th-cells. We have tested the modulating effect of L. lactis secreting human IL-10 (L.lacti s IL-10) on DC function in vitro. Monocyte-derived DC incubated wit

  19. Effect of X-Prolyl Dipeptidyl Aminopeptidase Deficiency on Lactococcus lactis

    NARCIS (Netherlands)

    Mayo, Baltasar; Kok, Jan; Bockelmann, Wilhelm; Haandrikman, Alfred; Leenhouts, Kees J.; Venema, Gerhardus

    1993-01-01

    The genetic determinant (pepXP) of an X-prolyl dipeptidyl aminopeptidase (PepXP) has recently been cloned and sequenced from both Lactococcus lactis subsp. cremoris (B. Mayo, J. Kok, K. Venema, W. Bockelmann, M. Teuber, H. Reinke, and G. Venema, Appl. Environ. Microbiol. 57:38-44, 1991) and L. lacti

  20. Complete genome sequence of the prototype lactic acid bacterium Lactococcus lactis subsp cremoris MG1363

    NARCIS (Netherlands)

    Wegmann, Udo; O'Connell-Motherwy, Mary; Zomer, Aldert; Buist, Girbe; Shearman, Claire; Canchaya, Carlos; Ventura, Marco; Goesmann, Alexander; Gasson, Michael J.; Kuipers, Oscar P.; van Sinderen, Douwe; Kok, Jan

    2007-01-01

    Lactococcus lactis is of great importance for the nutrition of hundreds of millions of people worldwide. This paper describes the genome sequence of Lactococcus lactis subsp. cremoris MG1363, the lactococcal strain most intensively studied throughout the world. The 2,529,478-bp genome contains 81 ps

  1. The Transcriptional and Gene Regulatory Network of Lactococcus lactis MG1363 during Growth in Milk

    DEFF Research Database (Denmark)

    de Jong, Anne; Hansen, Morten Ejby; Kuipers, Oscar P.;

    2013-01-01

    analysis of gene expression over time showed that L. lactis adapted quickly to the environmental changes. Using upstream sequences of genes with correlated gene expression profiles, we uncovered a substantial number of putative DNA binding motifs that may be relevant for L. lactis fermentative growth...

  2. A genome-based identification approach for members of the genus Bifidobacterium.

    Science.gov (United States)

    Ferrario, Chiara; Milani, Christian; Mancabelli, Leonardo; Lugli, Gabriele Andrea; Turroni, Francesca; Duranti, Sabrina; Mangifesta, Marta; Viappiani, Alice; Sinderen, Douwe van; Ventura, Marco

    2015-03-01

    During recent years, the significant and increasing interest in novel bifidobacterial strains with health-promoting characteristics has catalyzed the development of methods for efficient and reliable identification of Bifidobacterium strains at (sub) species level. We developed an assay based on recently acquired bifidobacterial genomic data and involving 98 primer pairs, called the Bifidobacterium-ampliseq panel. This panel includes multiplex PCR primers that target both core and variable genes of the pangenome of this genus. Our results demonstrate that the employment of the Bifidobacterium-ampliseq panel allows rapid and specific identification of the so far recognized 48 (sub)species harboring the Bifidobacterium genus, and thus represents a cost- and time-effective bifidobacterial screening methodology.

  3. Inhibitory activity of Bifidobacterium adolescent combined with cisplatin on melanoma in mice and its mechanism

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The aim of this study is to explore inhibitory activity of Bifidobacterium adolescent combined with cisplatin on the growth of melanoma(B16)in mice and the underlying mechanism.C57 mice were inoculated with B16 cancer cells to construct mouse model of melanoma and treated with bifidobacterium adolescent combined with cisplatin.Ratios of inhibitory activity on the growth of melanoma(B 16)were calculated.Pathology changes of the tumor were observed by HE staining.B 16 cell cycles were examined on a flow cytometer.Lymphocyte proliferation was measured with MTT assay and the T-cell subset was measured by double marked fluorescence.When bifidobacterium of 1010 cfu/L was injected,the ratio of inhibitory activity on the growth of melanoma(B16)reached 54%,which was similar to that of cisplatin group.The ratio of inhibitory activity reached 74.45% when the mice were treated by bifidobacterium combined with cisplatin,HE staining shows that bifidobacterium inhibited B16 cell proliferation and enhanced the cisplati(n)s killing activity on B16 cells.The results of flow cytometry demonstrated that B16 cell proliferation was arrested at G1 stage after treatment with bifidobacterium.The B16 cell proliferation was arrested at S stage after treatment with cisplatin.The CD4+ percentage increased and the difference was significant compared with the normal group after treatment with bifidobacterium,indicating that T-cell immune activity was enhanced.Treatment with bifidobacterium combined with cisplatin can enhance the inhibitory activity on the growth of melanoma(B16)of cisplatin.The mechanism of the inhibitory activity on B 16 cell proliferation is correlated with the enhanced immune activity in mice.

  4. PURIFICATION AND CHARACTERIZATION OF NISIN PRODUCED BY LACTOCOCCUS LACTIS ISOLATED FROM INDIAN CURD

    Directory of Open Access Journals (Sweden)

    Saba A. Mahdy

    2015-12-01

    Full Text Available Lactococcus lactis isolated from traditional dairy Indian curd. Strains were preliminarily identified by PCR analysis and partial 16S rRNA confirmed that N5 were 100% identical to Lactococcus. lactis sp. lactis. The results revealed that only the bacteriocin produced from strain N5 was shown as being active against mostly gram positive bacteria The bacteriocin produced purified by precipitation followed by loading with gel chromatography. The partially purified bacteriocin was found to be stable over a wide range of pH, temperature and enzymes. The molecular weight of the peptide was judged to be 3.5 kDa by SDSpolyacrylamide gel electrophoresis.and conform to the result of mass spectrometry by maldi-tof test which calculated the mass of 3354.07 Da for nisin.These results indicate that bacteriocin produced by L. lactis sp. lactis N5 is a nisin.

  5. Antimicrobial susceptibilities and random amplified polymorphic DNA-PCR fingerprint characterization of Lactococcus lactis ssp. lactis and Lactococcus garvieae isolated from bovine intramammary infections.

    Science.gov (United States)

    Plumed-Ferrer, C; Barberio, A; Franklin-Guild, R; Werner, B; McDonough, P; Bennett, J; Gioia, G; Rota, N; Welcome, F; Nydam, D V; Moroni, P

    2015-09-01

    In total, 181 streptococci-like bacteria isolated from intramammary infections (IMI) were submitted by a veterinary clinic to Quality Milk Production Services (QMPS, Cornell University, Ithaca, NY). The isolates were characterized by sequence analysis, and 46 Lactococcus lactis ssp. lactis and 47 Lactococcus garvieae were tested for susceptibility to 17 antibiotics. No resistant strains were found for β-lactam antibiotics widely used in clinical practice (penicillin, ampicillin, and amoxicillin), and all minimum inhibitory concentrations (MIC) were far from the resistance breakpoints. Eight strains had MIC intermediate to cefazolin. The random amplification of polymorphic DNA (RAPD)-PCR fingerprint patterns showed a slightly higher heterogeneity for Lc. lactis ssp. lactis isolates than for Lc. garvieae isolates.

  6. Partial characterization of an rpoD-like gene of Lactococcus lactis subsp. lactis ML3 with a polymerase chain reaction-based approach.

    Science.gov (United States)

    Gansel, X; Dutreix, M; Hartke, A; Boutibonnes, P; Auffray, Y

    1993-11-01

    With degenerated oligonucleotide primers for conserved regions of bacterial sigma factor proteins, a 117-bp internal DNA fragment of an rpoD-like gene of Lactococcus lactis subsp. lactis ML3 was amplified by the polymerase chain reaction (PCR). The DNA sequence of this PCR product was determined by cycle sequencing, and the deduced amino acid sequence of this internal fragment showed an extensive homology with the known sigma factor sequences from six other microorganisms and present a 13-amino acid region corresponding to the typical "RpoD box" of primary sigma factors. This PCR product was used as a probe to specifically detect sigma homologs in Pediococcus acidilactici, Leuconostoc lactis, Lactobacillus helveticus, Lactobacillus acidophilus, Enterococcus faecalis, Streptococcus thermophilus, and Lactococcus lactis subsp. cremoris. These data are consistent with the existence of a high similarity between the primary sigma factors from diverse Gram-positive microorganisms.

  7. Bifidobacterium in the gut microbiota confer resilience to chronic social defeat stress in mice.

    Science.gov (United States)

    Yang, Chun; Fujita, Yuko; Ren, Qian; Ma, Min; Dong, Chao; Hashimoto, Kenji

    2017-04-03

    Accumulating evidence suggests that abnormalities in the composition of the gut microbiota may play a role in the pathogenesis of depression. Although approximately 30% mice are resilient to chronic social defeat stress (CSDS), the role of gut microbiota in this stress resilience is unknown. In this study, male C57BL/6 mice were exposed to a different CD1 aggressor mouse for 10 min on 10 consecutive days. A social interaction test was applied to distinguish between resilient and susceptible mice. Using 16S rRNA analysis, we examined the composition of gut microbiota in feces from control, resilient, and susceptible mice. The marked appearance of Bifidobacterium was detected in the resilient mice, whereas in the control and susceptible mice, Bifidobacterium were below the detection limit. Oral intake of Bifidobacterium significantly increased the number of resilient mice after CSDS compared with vehicle-treated mice. These findings suggest that Bifidobacterium may confer resilience to CSDS. Therefore, supplementation of Bifidobacterium may prevent the onset of depression from stress in humans. In addition, supplementation of Bifidobacterium may prevent or minimize relapse from remission induced by inflammation and/or stress in depressed patients.

  8. Bifidobacterium in the gut microbiota confer resilience to chronic social defeat stress in mice

    Science.gov (United States)

    Yang, Chun; Fujita, Yuko; Ren, Qian; Ma, Min; Dong, Chao; Hashimoto, Kenji

    2017-01-01

    Accumulating evidence suggests that abnormalities in the composition of the gut microbiota may play a role in the pathogenesis of depression. Although approximately 30% mice are resilient to chronic social defeat stress (CSDS), the role of gut microbiota in this stress resilience is unknown. In this study, male C57BL/6 mice were exposed to a different CD1 aggressor mouse for 10 min on 10 consecutive days. A social interaction test was applied to distinguish between resilient and susceptible mice. Using 16S rRNA analysis, we examined the composition of gut microbiota in feces from control, resilient, and susceptible mice. The marked appearance of Bifidobacterium was detected in the resilient mice, whereas in the control and susceptible mice, Bifidobacterium were below the detection limit. Oral intake of Bifidobacterium significantly increased the number of resilient mice after CSDS compared with vehicle-treated mice. These findings suggest that Bifidobacterium may confer resilience to CSDS. Therefore, supplementation of Bifidobacterium may prevent the onset of depression from stress in humans. In addition, supplementation of Bifidobacterium may prevent or minimize relapse from remission induced by inflammation and/or stress in depressed patients. PMID:28368029

  9. Strain-Dependent Transcriptome Signatures for Robustness in Lactococcus lactis

    Science.gov (United States)

    Dijkstra, Annereinou R.; Alkema, Wynand; Starrenburg, Marjo J. C.; van Hijum, Sacha A. F. T.; Bron, Peter A.

    2016-01-01

    Recently, we demonstrated that fermentation conditions have a strong impact on subsequent survival of Lactococcus lactis strain MG1363 during heat and oxidative stress, two important parameters during spray drying. Moreover, employment of a transcriptome-phenotype matching approach revealed groups of genes associated with robustness towards heat and/or oxidative stress. To investigate if other strains have similar or distinct transcriptome signatures for robustness, we applied an identical transcriptome-robustness phenotype matching approach on the L. lactis strains IL1403, KF147 and SK11, which have previously been demonstrated to display highly diverse robustness phenotypes. These strains were subjected to an identical fermentation regime as was performed earlier for strain MG1363 and consisted of twelve conditions, varying in the level of salt and/or oxygen, as well as fermentation temperature and pH. In the exponential phase of growth, cells were harvested for transcriptome analysis and assessment of heat and oxidative stress survival phenotypes. The variation in fermentation conditions resulted in differences in heat and oxidative stress survival of up to five 10-log units. Effects of the fermentation conditions on stress survival of the L. lactis strains were typically strain-dependent, although the fermentation conditions had mainly similar effects on the growth characteristics of the different strains. By association of the transcriptomes and robustness phenotypes highly strain-specific transcriptome signatures for robustness towards heat and oxidative stress were identified, indicating that multiple mechanisms exist to increase robustness and, as a consequence, robustness of each strain requires individual optimization. However, a relatively small overlap in the transcriptome responses of the strains was also identified and this generic transcriptome signature included genes previously associated with stress (ctsR and lplL) and novel genes, including nan

  10. CTP limitation increases expression of CTP synthase in Lactococcus lactis

    DEFF Research Database (Denmark)

    Jørgensen, C.M.; Hammer, Karin; Martinussen, Jan

    2003-01-01

    for regulation of the pyrG gene. It is possible to fold the pyrG leader in an alternative structure that would prevent the formation of the terminator. We suggest a model for pyrG regulation in L. lactis, and probably in other gram-positive bacteria as well, in which pyrG expression is directly dependent...... on the CTP concentration through an attenuator mechanism. At normal CTP concentrations a terminator is preferentially formed in the pyrG leader, thereby reducing expression of CTP synthase. At low CTP concentrations the RNA polymerase pauses at a stretch of C residues in the pyrG leader, thereby allowing...

  11. Inhibition of Clostridium tyrobutyricum in Vidiago cheese by Lactococcus lactis ssp. lactis IPLA 729, a nisin Z producer.

    Science.gov (United States)

    Rilla, Natalia; Martínez, Beatriz; Delgado, Teresa; Rodríguez, Ana

    2003-08-15

    Lactococcus lactis ssp. lactis IPLA 729 is a nisin Z producer isolated from raw milk cheese able to grow and produce nisin Z in milk. The ability of this strain to inhibit the growth of Clostridium tyrobutyricum CECT 4011, a late blowing agent, in Vidiago cheese, a semi-hard farmhouse variety, manufactured in Asturias, Northern Spain, was investigated. For control purposes, cheeses were manufactured with the mesophilic mixed starter IPLA-001. In experimental cheeses, the nisin-producing strain L. lactis IPLA 729 was combined with this starter. Nisin Z activity reached a concentration of 1600 AU/ml in 1-day cheeses and this level was maintained until 15 days of ripening. Furthermore, to compare the inhibitory activity of the nisin-producing strain to nitrate, cheeses were also manufactured with a commercial starter culture and potassium nitrate as anti-blowing agent was added in accordance with Vidiago's cheesemakers. The control, experimental and commercial cheeses were contaminated with C. tyrobutyricum CECT 4011. The composition of the three different cheeses showed only slight differences with respect to total solids, protein and fat, although control and experimental cheeses showed a richer flavour-compound profile than commercial cheeses. The level of the spoilage strain C. tyrobutyricum CECT 4011 decreased from 1.2x10(6) to 1.3x10(3) cfu/g during ripening in presence of the nisin Z producer, while it increased to 1.99x10(9) cfu/g in control cheeses and to 3.5x10(7) cfu/g in commercial cheeses.

  12. Sequencing and transcriptional analysis of the biosynthesis gene cluster of putrescine-producing Lactococcus lactis.

    Science.gov (United States)

    Ladero, Victor; Rattray, Fergal P; Mayo, Baltasar; Martín, María Cruz; Fernández, María; Alvarez, Miguel A

    2011-09-01

    Lactococcus lactis is a prokaryotic microorganism with great importance as a culture starter and has become the model species among the lactic acid bacteria. The long and safe history of use of L. lactis in dairy fermentations has resulted in the classification of this species as GRAS (General Regarded As Safe) or QPS (Qualified Presumption of Safety). However, our group has identified several strains of L. lactis subsp. lactis and L. lactis subsp. cremoris that are able to produce putrescine from agmatine via the agmatine deiminase (AGDI) pathway. Putrescine is a biogenic amine that confers undesirable flavor characteristics and may even have toxic effects. The AGDI cluster of L. lactis is composed of a putative regulatory gene, aguR, followed by the genes (aguB, aguD, aguA, and aguC) encoding the catabolic enzymes. These genes are transcribed as an operon that is induced in the presence of agmatine. In some strains, an insertion (IS) element interrupts the transcription of the cluster, which results in a non-putrescine-producing phenotype. Based on this knowledge, a PCR-based test was developed in order to differentiate nonproducing L. lactis strains from those with a functional AGDI cluster. The analysis of the AGDI cluster and their flanking regions revealed that the capacity to produce putrescine via the AGDI pathway could be a specific characteristic that was lost during the adaptation to the milk environment by a process of reductive genome evolution.

  13. Genetically Modified Lactococcus lactis for Delivery of Human Interleukin-10 to Dendritic Cells

    Directory of Open Access Journals (Sweden)

    Inge L. Huibregtse

    2012-01-01

    Full Text Available Interleukin-10 (IL-10 plays an indispensable role in mucosal tolerance by programming dendritic cells (DCs to induce suppressor Th-cells. We have tested the modulating effect of L. lactis secreting human IL-10 (L.  lactisIL-10 on DC function in vitro. Monocyte-derived DC incubated with L.  lactisIL-10 induced effector Th-cells that markedly suppressed the proliferation of allogenic Th-cells as compared to L. lactis. This suppressive effect was only seen when DC showed increased CD83 and CD86 expression. Furthermore, enhanced production of IL-10 was measured in both L.  lactisIL-10-derived DC and Th-cells compared to L. lactis-derived DC and Th-cells. Neutralizing IL-10 during DC-Th-cell interaction and coculturing L.  lactisIL-10-derived suppressor Th-cells with allogenic Th-cells in a transwell system prevented the induction of suppressor Th-cells. Only 130 pg/mL of bacterial-derived IL-10 and 40 times more exogenously added recombinant human IL-10 were needed during DC priming for the generation of suppressor Th-cells. The spatially restricted delivery of IL-10 by food-grade bacteria is a promising strategy to induce suppressor Th-cells in vivo and to treat inflammatory diseases.

  14. In vitro evaluation of gastrointestinal survival of Lactobacillus amylovorus DSM 16698 alone and combined with galactooligosaccharides, milk and/or Bifidobacterium animalis subsp. lactis Bb-12

    NARCIS (Netherlands)

    Martinez, R.C.R.; Anynaou, A.E.; Albrecht, S.A.; Schols, H.A.; Martinis, de E.C.P.; Zoetendal, E.G.; Venema, K.; Saad, S.M.I.; Smidt, H.

    2011-01-01

    Probiotic properties of Lactobacillus amylovorus DSM 16698 were previously demonstrated in piglets. Here, its potential as a human probiotic was studied in vitro, using the TIM-1 system, which is fully validated to simulate the human upper gastrointestinal tract. To evaluate the effect of the food m

  15. Anti-inflammatory metabolite production in the gut from the consumption of probiotic yogurt containing Bifidobacterium animalis subsp. lactis LKM512.

    Science.gov (United States)

    Matsumoto, Mitsuharu; Benno, Yoshimi

    2006-06-01

    There is little evidence for a relationship between probiotic metabolites and host cytokine production. We investigated in the present study the possibility that anti-inflammatory metabolites can be produced in the gut by LKM512 yogurt consumption by using murine macrophage-like J774.1 cells and extracts prepared from the feces of elderly volunteers. These volunteers' acute inflammation had been inhibited by LKM512 yogurt consumption in a previous test. The tumor necrosis factor (TNF)-alpha production elicited in J774.1 cells stimulated by lipopolysaccharide (LPS) and in the fecal extracts obtained during the period of LKM512 yogurt consumption was significantly decreased (pyogurt consumption contributed to suppressing the inflammatory cytokine produced by macrophages and that one of the anti-inflammatory metabolites in the fecal extracts was likely to have been a polyamine.

  16. Study Protocol Effect of the consumption of a fermented dairy product containing Bifidobacterium lactis DN-173 010 on constipation in childhood: a multicentre randomised controlled trial (NTRTC: 1571)

    NARCIS (Netherlands)

    Tabbers, M.M.; Chmielewska, A.; Roseboom, M.G.; Boudet, C.; Perrin, C.; Szajewska, H.; Benninga, M.A.

    2009-01-01

    ABSTRACT: BACKGROUND: Constipation is a frustrating symptom affecting 3% of children worldwide. Randomised controlled trials show that both polyethylene glycol and lactulose are effective in increasing defecation frequency in children with constipation. However, in 30-50%, these children reported ab

  17. Probiotic treatment of collagenous colitis: a randomized, double-blind, placebo-controlled trial with Lactobacillus acidophilus and Bifidobacterium animalis subsp. Lactis

    DEFF Research Database (Denmark)

    Wildt, Signe; Munck, Lars K; Vinter-Jensen, Lars;

    2006-01-01

    Probiotic treatment may be effective in diseases involving gut microflora and intestinal inflammation. In collagenous colitis (CC), a potential pathogenic role of the gut microflora has been proposed. The effect of probiotic treatment in CC is unknown. Our aim was to investigate the clinical effect...

  18. Expression of Helicobacter pylori hspA Gene in Lactococcus lactis NICE System and Experimental Study on Its Immunoreactivity

    Directory of Open Access Journals (Sweden)

    Xiao-Juan Zhang

    2015-01-01

    Full Text Available Aim. The aim of this study was to develop an oral Lactococcus lactis (L. lactis vaccine against Helicobacter pylori (H. pylori. Methods. After L. lactis NZ3900/pNZ8110-hspA was constructed, growth curves were plotted to study whether the growth of recombinant L. lactis was affected after hspA was cloned into L. lactis and whether the growth of empty bacteria, empty plasmid bacteria, and recombinant L. lactis was affected by different concentrations of Nisin; SDS-PAGE and Western blot were adopted, respectively, to detect the HspA expressed by recombinant L. lactis and its immunoreactivity. Results. There was no effect observed from the growth curve after exogenous gene hspA was cloned into L. lactis NZ3900; different concentrations of Nisin did not affect the growth of NZ3900 and NZ3900/pNZ8110, while different concentrations of Nisin inhibited the growth of NZ3900/pNZ8110-hspA except 10 ng/mL Nisin. No HspA strip was observed from SDS-PAGE. Western blot analysis showed that HspA expressed by recombinant bacteria had favorable immunoreactivity. Conclusion. The growth of recombinant L. lactis was suppressed even though a small amount of HspA had been induced to express. Therefore recombinant L. lactis only express HspA which was not suitable to be oral vaccine against Helicobacter pylori.

  19. Exploring amino acid auxotrophy in Bifidobacterium bifidum PRL2010

    Directory of Open Access Journals (Sweden)

    Chiara eFerrario

    2015-11-01

    Full Text Available The acquisition and assimilation strategies followed by members of the infant gut microbiota to retrieve nitrogen from the gut lumen are still largely unknown. In particular, no information on these metabolic processes is available regarding bifidobacteria, which are among the first microbial colonizers of the human intestine. Here, evaluation of amino acid auxotrophy and prototrophy of Bifidobacterium bifidum, with particular emphasis on B. bifidum strain PRL2010 (LMG S-28692, revealed a putative auxotrophy for cysteine. In addition, we hypothesized that cysteine plays a role in the oxidative stress response in B. bifidum. The use of glutathione as an alternative reduced sulfur compound did not alleviate cysteine auxotrophy of this strain, though it was shown to stimulate expression of the genes involved in cysteine biosynthesis, reminiscent of oxidative stress response. When PRL2010 was grown on a medium containing complex substrates, such as whey proteins or casein hydrolysate, we noticed a distinct growth-promoting effect of these compounds. Transcriptional analysis involving B. bifidum PRL2010 cultivated on whey proteins or casein hydrolysate revealed that the biosynthetic pathways for cysteine and methionine are modulated by the presence of casein hydrolysate. Such findings support the notion that certain complex substrates may act as potential prebiotics for bifidobacteria in their ecological niche.

  20. Feruloyl oligosaccharides stimulate the growth of Bifidobacterium bifidum.

    Science.gov (United States)

    Yuan, Xiaoping; Wang, Jing; Yao, Huiyuan

    2005-08-01

    Insoluble dietary fiber from wheat bran contains some feruloyl groups linked to the arabinose residues in the cell wall arabinoxylan. Treatment of wheat bran insoluble dietary fiber with xylanase from Bacillus subtilis yielded feruloyl oligosacchairdes, which were purified with Amberlite XAD-2. Saponification of the feruloyl oligosaccharides released ferulic acid and arabinoxylan oligosaccharides which consist of arabinose and xylose. The effect of the feruloyl oligosacchairdes on the growth of Bifidobacterium bifidum F-35 was investigated in vitro. The B. bifidum produced acid when cultivated anaerobically in TPY broth with 0.5% feruloyl oligosacchairdes as the carbohydrate source. The biomass yield of the B. bifidum increased with increasing the concentration of feruloyl oligosaccharides in TPY broth. The maximum cell growth was increased by 50% in TPY broth supplemented with 0.1% feruloyl oligosaccharides compared to TPY broth. These results indicated that the growth of B. bifidum F-35 was promoted by the feruloyl oligosaccharides from wheat bran insoluble dietary fiber, and not suppressed by the ferulic acid moiety of them.

  1. Antioxidative properties and inhibitory effect of Bifidobacterium adolescentis on melanogenesis.

    Science.gov (United States)

    Huang, Huey-Chun; Chang, Tsong-Min

    2012-09-01

    Melanin is a dark pigment produced by melanocytes. Tyrosinase is a key enzyme which catalyzes the rate-limiting step of melanogenesis. However, accumulation of melanin leads to various skin hyperpigmentation disorders. To find a novel skin-whitening agent, the antioxidant capacity of Bifidobacterium adolescentis culture filtrate and inhibitory effect on melanogenesis were investigated. The antioxidant effects of B. adolescentis culture filtrate include 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging capacity, 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) radical cation scavenging activity and reducing power were measured spectrophotometrically. The reducing power is a useful index for the evaluation of potential antioxidants which carry out reduction of ferricyanide to ferrocyanide. Furthermore, the inhibitory effects of the bacterial culture filtrate on mushroom tyrosinase, B16F10 intracellular tyrosinase activity and melanin content were also determined. The results revealed that B. adolescentis culture filtrate (2.5, 5.0 and 7.5 %; v/v) effectively scavenged DPPH and ABTS radicals, and lower concentrations of the bacterial culture filtrates (0.5, 1.0 and 1.5 %; v/v) showed potent reducing power in a dose-dependent pattern. Additionally, the bacterial culture filtrate suppressed murine tyrosinase activity and decreased the amount of melanin in a dose-dependent manner. Our results demonstrated that B. adolescentis culture filtrate decreases the melanogenesis process of melanoma cells by inhibiting tyrosinase activity, which we suggest may be mediated through its antioxidant activity.

  2. An evaluation and partial characterization of a bacteriocin produced by Lactococcus lactis subsp lactis ST1 isolated from goat milk

    Directory of Open Access Journals (Sweden)

    Parinaz Taheri

    2012-12-01

    Full Text Available A bacteriocin-like inhibitory substance producing Lactococcus lactis subsp lactis strain, ST1, isolated from goat milk of Iranian origin and with broad spectrum of activity and desirable technical properties was used for evaluating some futures of bacteriocin inhibitory activity. Cell growth and bacteriocin production studies were carried out in MRS medium incubated statically under uncontrolled pH condition. The antibacterial activity presented a primary metabolite pattern and showed a rapid decrease at the stationary phase. Microaerobiosis and capnophily growth conditions resulted in higher bacteriocin production while aerobiosis showed negative effect on both cell growth and bacteriocin production. Bacteriocin production, on the other hand, was favored in MRS broth (pH; 6.5 inoculated with 0.1 ml l-1 fresh culture when incubation was carried out at 30 °C. This indicated that the conditions resulted in higher levels of growth were frequently favoring bacteriocin production by ST1 as well. Decrease in activity, at the stationary growth phase, was much pronounced in favored growth condition. Nutrient depletion, deferent effect of low pH on bacteriocin production and/or protein degradation seemed more responsible for this phenomenon. The study also provided further data on new method for bacteriocin release from the cell wall of producer. It was clearly shown that both heating and ultrasound shock for 5 min at pH 2 could increase bacteriocin activity significantly. The release was more pronounced in the presence of 0.5% Tween80.

  3. Antimicrobial activity of the Nisin Z producer Lactococcus lactis subsp. lactis Lc08 against Listeria monocytogenes in skim milk

    Directory of Open Access Journals (Sweden)

    L.M. Perin

    2013-10-01

    Full Text Available The presented study aimed to verify the effect of different pH values, enzyme solutions and heat treatments on the antimicrobial activity of the bacteriocinogenic strain Lactococcus lactis subsp. lactis Lc08 and to test their antimicrobial activity against Listeria monocytogenes in reconstituted skim milk at refrigeration temperatures. This strain was previously described as a nisin Z producer and capable of inhibiting L. monocytogenes growth in in vitro tests. The antimicrobial activity of the bacteriocin cell-free supernatant of Lc08 was sensitive to enzyme treatments (except papain. The pH values and heating (65ºC for 30min, 75ºC for 15s had no apparent effect on the antimicrobial activity of the bacteriocin produced by Lc08. Only treatment at autoclave conditions result in loss of their antimicrobial activity. Lc08 presented antimicrobial activity against L. monocytogenes in the milk system after 12h at 25ºC. No effect was found at 7ºC. The results show the application viability of the Lc08 in food systems as a biopreservative against L. monocytogenes.

  4. L+-lactic acid production from starch by a novel amylolytic Lactococcus lactis subsp. lactis B84.

    Science.gov (United States)

    Petrov, Kaloyan; Urshev, Zoltan; Petrova, Penka

    2008-06-01

    A new Lactococcus lactis subsp. lactis B84, capable of utilizing starch as a sole carbon source and producing L(+)-lactate, was isolated from spontaneously fermented rye sourdough. Aiming at maximum lactic acid productivity, the components of the media and the cultivation conditions were varied. In MRS-starch medium (with absence of yeast and meat extracts), at 33 degrees C, agitation 200 rpm and pH 6.0 for 6 days complete starch hydrolysis occurred and 5.5 gl(-1) lactic acid were produced from 18 gl(-1) starch. The identification of strain B84 was based on genetic criteria. Amplified ribosomal DNA restriction analysis (ARDRA), PCR with species-specific primers and sequencing of the 16S rDNA proved its species affiliation. Four genes for enzymes, involved in starch degradation were detected in B84 genome: amyL, amyY, glgP and apu, coding cytoplasmic and extracellular alpha-amylases, glycogen phosphorylase and amylopullulanase, respectively. Reverse transcription PCR experiments showed that both genes, encoding alpha-amylases (amyL and amyY) were expressed into mRNAs, whereas apu and glgP were not. Amylase activity assay was performed at different pH and temperatures. The cell-bond amylase proved to be the key enzyme, involved in the starch hydrolysis with maximum activity at 45 degrees C and pH 5.4.

  5. Localization and accessibility of antigenic sites of the extracellular serine proteinase of Lactococcus lactis

    NARCIS (Netherlands)

    Laan, Harm; Kok, Jan; Haandrikman, Alfred J.; Venema, Gerhardus; Konings, Wilhelmus

    1992-01-01

    Lactococcus lactis strains produce an extracellular subtilisin-related serine proteinase in which immunologically different components can be distinguished. Monoclonal antibodies specific for the different proteinase components have been raised and their epitopes were identified. By Western-blot ana

  6. Physiological Adaptation of the Bacterium Lactococcus lactis in Response to the Production of Human CFTR

    NARCIS (Netherlands)

    Steen, Anton; Wiederhold, Elena; Gandhi, Tejas; Breitling, Rainer; Slotboom, Dirk Jan

    2011-01-01

    Biochemical and biophysical characterization of CFTR (the cystic fibrosis transmembrane conductance regulator) is thwarted by difficulties to obtain sufficient quantities of correctly folded and functional protein. Here we have produced human CFTR in the prokaryotic expression host Lactococcus lacti

  7. Bifidobacterium asteroides PRL2011 genome analysis reveals clues for colonization of the insect gut.

    Directory of Open Access Journals (Sweden)

    Francesca Bottacini

    Full Text Available Bifidobacteria are known as anaerobic/microaerophilic and fermentative microorganisms, which commonly inhabit the gastrointestinal tract of various animals and insects. Analysis of the 2,167,301 bp genome of Bifidobacterium asteroides PRL2011, a strain isolated from the hindgut of Apis mellifera var. ligustica, commonly known as the honey bee, revealed its predicted capability for respiratory metabolism. Conservation of the latter gene clusters in various B. asteroides strains enforces the notion that respiration is a common metabolic feature of this ancient bifidobacterial species, which has been lost in currently known mammal-derived Bifidobacterium species. In fact, phylogenomic based analyses suggested an ancient origin of B. asteroides and indicates it as an ancestor of the genus Bifidobacterium. Furthermore, the B. asteroides PRL2011 genome encodes various enzymes for coping with toxic products that arise as a result of oxygen-mediated respiration.

  8. Bifidobacterium asteroides PRL2011 genome analysis reveals clues for colonization of the insect gut.

    Science.gov (United States)

    Bottacini, Francesca; Milani, Christian; Turroni, Francesca; Sánchez, Borja; Foroni, Elena; Duranti, Sabrina; Serafini, Fausta; Viappiani, Alice; Strati, Francesco; Ferrarini, Alberto; Delledonne, Massimo; Henrissat, Bernard; Coutinho, Pedro; Fitzgerald, Gerald F; Margolles, Abelardo; van Sinderen, Douwe; Ventura, Marco

    2012-01-01

    Bifidobacteria are known as anaerobic/microaerophilic and fermentative microorganisms, which commonly inhabit the gastrointestinal tract of various animals and insects. Analysis of the 2,167,301 bp genome of Bifidobacterium asteroides PRL2011, a strain isolated from the hindgut of Apis mellifera var. ligustica, commonly known as the honey bee, revealed its predicted capability for respiratory metabolism. Conservation of the latter gene clusters in various B. asteroides strains enforces the notion that respiration is a common metabolic feature of this ancient bifidobacterial species, which has been lost in currently known mammal-derived Bifidobacterium species. In fact, phylogenomic based analyses suggested an ancient origin of B. asteroides and indicates it as an ancestor of the genus Bifidobacterium. Furthermore, the B. asteroides PRL2011 genome encodes various enzymes for coping with toxic products that arise as a result of oxygen-mediated respiration.

  9. Lactococcus lactis metabolism and gene expression during growth on plant tissues.

    Science.gov (United States)

    Golomb, Benjamin L; Marco, Maria L

    2015-01-01

    Lactic acid bacteria have been isolated from living, harvested, and fermented plant materials; however, the adaptations these bacteria possess for growth on plant tissues are largely unknown. In this study, we investigated plant habitat-specific traits of Lactococcus lactis during growth in an Arabidopsis thaliana leaf tissue lysate (ATL). L. lactis KF147, a strain originally isolated from plants, exhibited a higher growth rate and reached 7.9-fold-greater cell densities during growth in ATL than the dairy-associated strain L. lactis IL1403. Transcriptome profiling (RNA-seq) of KF147 identified 853 induced and 264 repressed genes during growth in ATL compared to that in GM17 laboratory culture medium. Genes induced in ATL included those involved in the arginine deiminase pathway and a total of 140 carbohydrate transport and metabolism genes, many of which are involved in xylose, arabinose, cellobiose, and hemicellulose metabolism. The induction of those genes corresponded with L. lactis KF147 nutrient consumption and production of metabolic end products in ATL as measured by gas chromatography-time of flight mass spectrometry (GC-TOF/MS) untargeted metabolomic profiling. To assess the importance of specific plant-inducible genes for L. lactis growth in ATL, xylose metabolism was targeted for gene knockout mutagenesis. Wild-type L. lactis strain KF147 but not an xylA deletion mutant was able to grow using xylose as the sole carbon source. However, both strains grew to similarly high levels in ATL, indicating redundancy in L. lactis carbohydrate metabolism on plant tissues. These findings show that certain strains of L. lactis are well adapted for growth on plants and possess specific traits relevant for plant-based food, fuel, and feed fermentations.

  10. Efeito e modo de ação das bacteriocinas produzidas por Lactococcus lactis subsp. lactis ITAL 383, ATCC 11454 e CNRZ 150 contra Listeria innocua LIN 11 Effect and mode of action of the bacterioncin produced by Lactococcus. lactis subsp. lactis ITAL 383, ATCC 11454 e CNRZ 150 against Listeria innocua LIN 11

    Directory of Open Access Journals (Sweden)

    Izildinha MORENO

    1999-01-01

    Full Text Available O efeito e o modo de ação das bacteriocinas produzidas por L. lactis subsp. lactis ITAL 383 e CNRZ 150 são similares à nisina de L. lactis subsp. lactis ATCC 11454. Estas bacteriocinas apresentaram um modo de ação bactericida, causando a lise de células de L. innocua LIN 11, associada ao decréscimo da absorbância e da viabilidade celular. O efeito letal foi maior para células em fase exponencial comparativamente à fase estacionária de crescimento. A adsorção dessas bacteriocinas às células de L. innocua LIN 11 foi muito rápida e influenciada pelo pH do meio de suspensão; adsorção máxima foi verificada a pH 6,0 e logo após o contato inicial. Perda completa de adsorção ocorreu em pH 2,0.The effect and mode of action of the bacteriocin produced by L. lactis subsp. lactis ITAL 383 and CNRZ 150 are similar to the nisin produced by L. lactis subsp. lactis ATCC 11454. It was clearly bactericidal, and caused lysis of a strain of L. innocua LIN 11 detected by the decrease of absorbance values and the cell viability. Their lethal effect was considerably higher during the logarithmic growth when compared to the stationary phase. Adsorption developed rapidly and was influenced by the pH value of the suspension medium. Maximum adsorption was observed at pH 6,0 and immediately after initial contact and loss at pH 2,0.

  11. Fate of Lactococcus lactis starter cultures during late ripening in cheese models.

    Science.gov (United States)

    Ruggirello, Marianna; Cocolin, Luca; Dolci, Paola

    2016-10-01

    The presence of Lactococcus lactis, commonly employed as starter culture, was, recently, highlighted and investigated during late cheese ripening. Thus, the main goal of the present study was to assess the persistence and viability of this microorganism throughout manufacturing and ripening of model cheeses. Eight commercial starters, constituted of L. lactis subsp. lactis and L. lactis subsp. cremoris, were inoculated in pasteurized milk in order to manufacture miniature cheeses, ripened for six months. Samples were analysed at different steps (milk after inoculum, curd after cutting, curd after pressing and draining, cheese immediately after salting and cheese at 7, 15, 30, 60, 90, 120, 150 and 180 days of ripening) and submitted to both culture-dependent (traditional plating on M17) and -independent analysis (reverse transcription-quantitative PCR). On the basis of direct RNA analysis, L. lactis populations were detected in all miniature cheeses up to the sixth month of ripening, confirming the presence of viable cells during the whole ripening process, including late stages. Noteworthy, L. lactis was detected by RT-qPCR in cheese samples also when traditional plating failed to indicate its presence. This discrepancy could be explain with the fact that lactococci, during ripening process, enter in a stressed physiological state (viable not culturable, VNC), which might cause their inability to grow on synthetic medium despite their viability in cheese matrix. Preliminary results obtained by "resuscitation" assays corroborated this hypothesis and 2.5% glucose enrichment was effective to recover L. lactis cells in VNC state. The capability of L. lactis to persist in late ripening, and the presence of VNC cells which are known to shift their catabolism to peptides and amino acids consumption, suggests a possible technological role of this microorganism in cheese ripening with a possible impact on flavour formation.

  12. Over-expressed CmbT multidrug resistance transporter improves the fitness of Lactococcus lactis

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    Filipić Brankica

    2013-01-01

    Full Text Available The influence of the over-expression of CmbT multidrug resistance transporter on the growth rate of Lactococcus lactis NZ9000 was studied. L. lactis is a lactic acid bacteria (LAB widely used as a starter culture in dairy industry. Recently characterized CmbT MDR transporter in L. lactis confers resistance to a wide variety of toxic compounds as well as to some clinically relevant antibiotics. In this study, the cmbT gene was over-expressed in the strain L. lactis NZ9000 in the presence of nisin inducer. Over-expression of the cmbT gene in L. lactis NZ9000 was followed by RT-PCR. The obtained results showed that the cmbT gene was successfully over-expressed by addition of sub-inhibitory amounts of nisin. Growth curves of L. lactis NZ9000/pCT50 over-expressing the cmbT gene and L. lactis NZ9000 control strain were followed in the rich medium as well as in the chemically defined medium in the presence solely of methionine (0.084 mM or mix of methionine and cysteine (8.4 mM and 8.2 mM, respectively. Resulting doubling times revealed that L. lactis NZ9000/pCT50 had higher growth rate comparing to the control strain. This could be a consequence of the CmbT efflux activity, which improves the fitness of the host bacterium through the elimination of toxic compounds from the cell.

  13. Expanding the molecular toolbox for Lactococcus lactis: construction of an inducible thioredoxin gene fusion expression system

    LENUS (Irish Health Repository)

    Douillard, Francois P

    2011-08-09

    Abstract Background The development of the Nisin Inducible Controlled Expression (NICE) system in the food-grade bacterium Lactococcus lactis subsp. cremoris represents a cornerstone in the use of Gram-positive bacterial expression systems for biotechnological purposes. However, proteins that are subjected to such over-expression in L. lactis may suffer from improper folding, inclusion body formation and\\/or protein degradation, thereby significantly reducing the yield of soluble target protein. Although such drawbacks are not specific to L. lactis, no molecular tools have been developed to prevent or circumvent these recurrent problems of protein expression in L. lactis. Results Mimicking thioredoxin gene fusion systems available for E. coli, two nisin-inducible expression vectors were constructed to over-produce various proteins in L. lactis as thioredoxin fusion proteins. In this study, we demonstrate that our novel L. lactis fusion partner expression vectors allow high-level expression of soluble heterologous proteins Tuc2009 ORF40, Bbr_0140 and Tuc2009 BppU\\/BppL that were previously insoluble or not expressed using existing L. lactis expression vectors. Over-expressed proteins were subsequently purified by Ni-TED affinity chromatography. Intact heterologous proteins were detected by immunoblotting analyses. We also show that the thioredoxin moiety of the purified fusion protein was specifically and efficiently cleaved off by enterokinase treatment. Conclusions This study is the first description of a thioredoxin gene fusion expression system, purposely developed to circumvent problems associated with protein over-expression in L. lactis. It was shown to prevent protein insolubility and degradation, allowing sufficient production of soluble proteins for further structural and functional characterization.

  14. Determination of Bifidobacterium and Lactobacillus in breast milk of healthy women by digital PCR.

    Science.gov (United States)

    Qian, L; Song, H; Cai, W

    2016-09-01

    Breast milk is one of the most important sources of postnatal microbes. Quantitative real-time polymerase chain reaction (qRT-PCR) is currently used for the quantitative analysis of bacterial 16S rRNA genes in breast milk. However, this method relies on the use of standard curves and is imprecise when quantitating target DNA of low abundance. In contrast, droplet digital PCR (DD-PCR) provides an absolute quantitation without the need for calibration curves. A comparison between DD-PCR and qRT-PCR was conducted for the quantitation of Bifidobacterium and Lactobacillus 16S RNA genes in human breast milk, and the impacts of selected maternal factors were studied on the composition of these two bacteria in breast milk. From this study, DD-PCR reported between 0-34,460 16S rRNA gene copies of Bifidobacterium genera and between 1,108-634,000 16S rRNA gene copies of Lactobacillus genera in 1 ml breast milk. The 16S rRNA gene copy number of Lactobacillus genera was much greater than that of Bifidobacterium genera in breast milk. DD-PCR showed a 10-fold lower limit of quantitation as compared to qRT-PCR. A higher correlation and agreement was observed between qRT-PCR and DD-PCR in Lactobacillus quantitation as compared to Bifidobacterium quantitation. Based on our DD-PCR quantitation, a low abundance of Bifidobacterium bacteria in breast milk was correlated to higher pre-pregnancy body mass index (BMI). However, no significant difference was observed for these two bacteria in breast milk between mothers who had vaginal deliveries and caesarean deliveries. This study suggests that DD-PCR is a better tool to quantitate the bacterial load of breast milk compared to the conventional qRT-PCR method. The number of breast milk Bifidobacterium bacteria is influenced by maternal pre-pregnancy BMI.

  15. A probiotic treatment containing Lactobacillus, Bifidobacterium and Enterococcus improves IBS symptoms in an open label trial

    Institute of Scientific and Technical Information of China (English)

    FAN Yu-jing; CHEN Shu-jie; YU Ying-cong; SI Jian-min; LIU Bin

    2006-01-01

    Objective: To evaluate the efficacy and safety of live combined Bifidobacterium, Lactobacillus and Enterococcus capsules in treatment of irritable bowel syndrome. Methods: Eighty-five patients [male 32, female 53; age (45.31±11.72) years]were given live combined Bifidobacterium, Lactobacillus and Enterococcus capsules 1260 mg/d t.i.d.×4 weeks. Syndrome scales were used to evaluate the efficacy in gastrointestinal syndrome. Fecal flora was also measured before and after the treatment. Six bacteria were cultured and the colony forming units were counted in stool. SPSS was used for data analysis. Results: Seventy-four patients finished the follow-up. No side-effect was found. For treatment of irritable bowel syndrome, the effective rate of live combined Bifidobacterium, Lactobacillus and Enterococcus capsules was 56.8% in the second week, 74.3% in the fourth week and 73.0% in the sixth week. Single symptom was improved, especially in abdominal pain and stool character. The probiotica containing live combined Bifidobacterium, Lactobacillus and Enterococcus could increase bifidobacterium count (P<0.01) and lactobacillus count (P<0.05); decrease bacteroides count (P<0.05) and enterococci count (P<0.01); No obvious changes were observed in clostridium difficile colonitis and enterobacteriaceae (P>0.05). Conclusion: The result of the study indicated that the administration of live combined Bifidobacterium, Lactobacillus and Enterococcus improved the symptom of irritable bowel syndrome and that there was a gradual increase of this effect. Thereafter conditions remained stable for 2 weeks. That improvement may be associated with alterations in gastrointestinal flora.

  16. Differential expression of proteins and genes in the lag phase of Lactococcus lactis subsp lactis grown in synthetic medium and reconstituted skim milk

    DEFF Research Database (Denmark)

    Larsen, N.; Boye, Mette; Jakobsen, Marianne

    2006-01-01

    metabolism, glycolysis, stress response, translation, transcription, cell division, amino acid metabolism, and coenzyme synthesis., were identified. Among the identified proteins, > 2-fold induction and down-regulation in the lag phase were determined for 12 proteins in respect to the exponential phase......We investigated protein and gene expression in the lag phase of Lactococcus lactis subsp. lactis CNRZ 157 and compared it to the exponential and stationary phases. By means of two-dimensional polyacrylamide gel electrophoresis, 28 highly expressed lag-phase proteins, implicated in nucleotide...... and for 18 proteins in respect to the stationary phase. Transcriptional changes of the lag-phase proteins in L. lactis were studied by oligonucleotide microarrays. Good correlation between protein and gene expression studies was demonstrated for several differentially expressed proteins, including nucleotide...

  17. Electroinduced extraction of beta-galactosidase from Kluyveromyces lactis.

    Science.gov (United States)

    Ganeva, V; Galutzov, B; Eynard, N; Teissié, J

    2001-08-01

    A new methodology for the extraction of beta-galactosidase from the yeast Kluyveromyces lactis was obtained by electropulsation. The application of a series of electric pulses (2 ms duration, 1 Hz frequency, and 4-4.5 kV/cm field strength) to fresh cells suspended in deionized water, followed by incubation in PBS, led to a spontaneous slow release of enzyme at a yield of 75-80% without any further treatment. Most of the enzyme was extracted within 8 h after electropulsation. This release was dependent on the growth phase. The specific activity of beta-galactosidase in the supernatant of pulsed cells was higher by a factor of 1.5-1.7 in comparison with crude extract.

  18. Antigenicity and Immunogenicity of Rotavirus VP6 Protein Expressed on the Surface of Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    L. E. Esteban

    2013-01-01

    Full Text Available Group A rotaviruses are the major etiologic agents of acute gastroenteritis worldwide in children and young animals. Among its structural proteins, VP6 is the most immunogenic and is highly conserved within this group. Lactococcus lactis is a food-grade, Gram-positive, and nonpathogenic lactic acid bacteria that has already been explored as a mucosal delivery system of heterologous antigens. In this work, the nisin-controlled expression system was used to display the VP6 protein at the cell surface of L. lactis. Conditions for optimal gene expression were established by testing different nisin concentrations, cell density at induction, and incubation times after induction. Cytoplasmic and cell wall protein extracts were analyzed by Western blot and surface expression was confirmed by flow cytometry. Both analysis provided evidence that VP6 was efficiently expressed and displayed on the cell surface of L. lactis. Furthermore, the humoral response of mice immunized with recombinant L. lactis was evaluated and the displayed recombinant VP6 protein proved to be immunogenic. In conclusion, this is the first report of displaying VP6 protein on the surface of L. lactis to induce a specific immune response against rotavirus. These results provide the basis for further evaluation of this VP6-displaying L. lactis as a mucosal delivery vector in a mouse model of rotavirus infection.

  19. Genome shuffling of Lactococcus lactis subspecies lactis YF11 for improving nisin Z production and comparative analysis.

    Science.gov (United States)

    Zhang, Y F; Liu, S Y; Du, Y H; Feng, W J; Liu, J H; Qiao, J J

    2014-05-01

    Nisin has been widely used in the food industry as a safe and natural preservative to increase the shelf time of many foods. In this study, genome shuffling was applied to improve nisin Z production of Lactococcus lactis ssp. lactis YF11 (YF11) via recursive protoplast fusion. Ultraviolet irradiation and diethyl sulfate mutagenesis were used to generate parental strains for genome shuffling. After 4 rounds of genome shuffling, the best-performing strain F44 was obtained, which showed dramatic improvements in tolerance to both glucose (ranging from 8 to 15% (wt/vol) and nisin (ranging from 5,000 to 14,000 IU/mL). Fed-batch fermentation showed that the nisin titer of F44 was up to 4,023 IU/mL, which was 2.4 times that of the starting strain YF11. Field emission scanning electron microscope micrographs of YF11 and F44 revealed the apparent differences in cell morphology. Whereas YF11 displayed long and thin cell morphology, F44 cells were short and thick and with a raised surface in the middle of the cell. With the increasing glucose and nisin content in the medium, cells of both YF11 and F44 tended to become shrunken; however, alterations in YF11 cells were more pronounced than those of F44 cells, especially when cultured in tolerance medium containing both nisin and glucose. Nuclear magnetic resonance analysis demonstrated that the structure of nisin from YF11 and F44 was the same. Expression profiling of nisin synthesis related genes by real-time quantitative PCR showed that the transcription level of nisin structural gene nisZ and immunity gene nisI of F44 was 48 and 130% higher than that of the starting strain YF11, respectively. These results could provide valuable insights into the molecular basis underlying the nisin overproduction mechanism in L. lactis, thus facilitating the future construction of industrial strains for nisin production.

  20. Glycosylation in secreted proteins from yeast Kluyveromyces lactis

    Energy Technology Data Exchange (ETDEWEB)

    Santos, A.V.; Passos, F.M.L. [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Microbiologia. Lab. de Fisiologia de Microrganismos; Azevedo, B.R.; Pimenta, A.M.C.; Santoro, M.M. [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Dept. de Bioquimica e Imunologia. Lab. de Enzimologia e Fisico-Quimica de Proteina

    2008-07-01

    Full text: The nutritional status of a cell culture affects either the expression or the traffic of a number of proteins. The identification of the physiological conditions which favor protein secretion has important biotechnological consequences in designing systems for recombinant extracellular protein industrial production. Yeast Kluyvromyces lactis has been cultured in a continuous stirring tank bioreactor (CSTR) under nitrogen limitation at growth rates (0.03 h{sup -1} and 0.09 h{sup -1}) close to either exponential or stationary batch growth phases, respectively the objective was to investigate the extracellular glycoproteins at these two level of nitrogen limitation. Proteins from free cell extracts were separated by gradient SDS-PAGE (5-15%) and two-dimensional chromatography, and were analyzed by mass spectrometry (MALDI-TOF-TOF-MS). In SDS-PAGE analysis, differences in extracellular proteome were visualized: different proteins profiles at these two growth rates. The 0.09 h-1 growth rate showed larger number of bands using colloidal Coma ssie Blue staining. Different bands were detected at these two growth rates when the PAS assay for glycoprotein detection in polyacrylamide gel was used. The two-dimensional chromatogram profiles were comparatively distinguished between the 0.03 h{sup -1} and 0.09 h{sup -1} growth rate samples. Protein peaks from the second dimension, were subjected to mass spectrometry. The mass spectrums visualized showed glycosylated proteins with N-acetylglucosamine molecules and 8, 9 or 15 hexoses molecules. Comparisons between the proteins averaged mass values with the deduced proteins masses from K. lactis secreted proteins database indicated possible post-translational modifications, such as post-translational proteolysis, acetylation, deamidation and myristoylation.

  1. Viability of Two Freeze-dried Strains of Bifidobacterium Preparations at Various Temperatures during Prolonged Storage

    Institute of Scientific and Technical Information of China (English)

    DU Peng; DU Li-hui; HUO Gui-cheng

    2005-01-01

    Viability of bifidobacteria in freeze-dried probiotic products at various temperatures during prolonged storage was assessed. Bifidobacterium longum and Bifidobacterium infantis were freeze-dried. The freeze-dried preparations were stored at -18,4, and 20℃. Cell counts were enumerated using BS agar at 37℃ for 48 h under anaerobic conditions at 0, 45 and 120 days. Storage at 20℃ showed the greatest decline in the viability of bifidobacteria, whereas that at -18℃ showed the least decrease.

  2. Bifidobacterium animalis AHC7 protects against pathogen-induced NF-kappaB activation in vivo

    LENUS (Irish Health Repository)

    O'Mahony, David

    2010-12-22

    Abstract Background Bifidobacteria and lactobacilli are among the early and important colonizers of the gastrointestinal tract and are generally considered to be part of a normal, healthy microbiota. It is believed that specific strains within the microbiota can influence host immune-reactivity and may play a role in protection from infection and aberrant inflammatory activity. One such strain, Bifidobacterium animalis AHC7, has been previously shown to protect against Salmonella typhimurium infection in mice and helps resolve acute idiopathic diarrhea in dogs. The aim of this study was to investigate the potential molecular and cellular mechanisms underpinning the Bifidobacterium animalis AHC7 protective effect. Results Following 4 hours of infection with Salmonella typhimurium, NF-κB activation was significantly elevated in vivo in placebo and Enterococcus faecium-fed animals while Bifidobacterium animalis AHC7 consumption significantly attenuated the NF-κB response. In vitro anti-CD3\\/CD28 stimulated Peyer\\'s patch cells secreted significantly less TNF-α and IFN-γ following Bifidobacterium animalis AHC7 consumption. Stimulated cells released more IL-12p70 but this difference did not reach statistical significance. No alteration in mucosal IL-6, IL-10 or MCP-1 levels were observed. No statistically significant change in the cytokine profile of mesenteric lymph node cells was noted. In vitro, Bifidobacterium animalis AHC7 was bound by dendritic cells and induced secretion of both IL-10 and IL-12p70. In addition, co-culture of CD4+ T cells with Bifidobacterium animalis AHC7-stimulated dendritic cells resulted in a significant increase in CD25+Foxp3+ T cell numbers. Conclusion Bifidobacterium animalis AHC7 exerts an anti-inflammatory effect via the attenuation of pro-inflammatory transcription factor activation in response to an infectious insult associated with modulation of pro-inflammatory cytokine production within the mucosa. The cellular mechanism

  3. Bifidobacterium animalis AHC7 protects against pathogen-induced NF-κB activation in vivo

    Directory of Open Access Journals (Sweden)

    Scully Paul

    2010-12-01

    Full Text Available Abstract Background Bifidobacteria and lactobacilli are among the early and important colonizers of the gastrointestinal tract and are generally considered to be part of a normal, healthy microbiota. It is believed that specific strains within the microbiota can influence host immune-reactivity and may play a role in protection from infection and aberrant inflammatory activity. One such strain, Bifidobacterium animalis AHC7, has been previously shown to protect against Salmonella typhimurium infection in mice and helps resolve acute idiopathic diarrhea in dogs. The aim of this study was to investigate the potential molecular and cellular mechanisms underpinning the Bifidobacterium animalis AHC7 protective effect. Results Following 4 hours of infection with Salmonella typhimurium, NF-κB activation was significantly elevated in vivo in placebo and Enterococcus faecium-fed animals while Bifidobacterium animalis AHC7 consumption significantly attenuated the NF-κB response. In vitro anti-CD3/CD28 stimulated Peyer's patch cells secreted significantly less TNF-α and IFN-γ following Bifidobacterium animalis AHC7 consumption. Stimulated cells released more IL-12p70 but this difference did not reach statistical significance. No alteration in mucosal IL-6, IL-10 or MCP-1 levels were observed. No statistically significant change in the cytokine profile of mesenteric lymph node cells was noted. In vitro, Bifidobacterium animalis AHC7 was bound by dendritic cells and induced secretion of both IL-10 and IL-12p70. In addition, co-culture of CD4+ T cells with Bifidobacterium animalis AHC7-stimulated dendritic cells resulted in a significant increase in CD25+Foxp3+ T cell numbers. Conclusion Bifidobacterium animalis AHC7 exerts an anti-inflammatory effect via the attenuation of pro-inflammatory transcription factor activation in response to an infectious insult associated with modulation of pro-inflammatory cytokine production within the mucosa. The cellular

  4. New Antifungal Bacteriocin-Synthesizing Strains of Lactococcus lactis ssp. lactis as the Perspective Biopreservatives for Protection of Raw Smoked Sausages

    Directory of Open Access Journals (Sweden)

    L. G. Stoyanova

    2010-01-01

    Full Text Available Problem statement: Screening for the effective bacteriocin-synthesizing strains of Lactococcus lactis as the perspective biopreservatives was performed. We used a raw milk and dairy products from different climatic regions as well as from powerful drinks of mixed lactic acid and alcoholic fermentation: kurunga, kumiss and Iranian Dough, that were widely used by local population to prevent diseases. Approach: The special interest was paid to isolates of lactococci with antagonistic activity. According to their morphological, cultural, physiological, biochemical properties and sequence of 16S rRNA gene they were identified as Lactococcus lactis ssp. lactis. Only nine from the selected 94 strains expressed a broad spectrum of activity against Gram-positive and Gramnegative bacteria including pathogens (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris, Salmonella gallinarum, moulds (Aspergillus, Fusarium, Penicillium genera, as well as yeasts (Rhodotorula, Candida. Results: It reveals the unique biological properties for isolated natural strains of Lactococcus lactis species. Most effective new bacteriocin-synthesizing strains 194 and K-205 were isolated from raw cow milk and kurunga from Buryatia. These strains had high antibiotic activity up to 3600 and 2700 IU mL-1 as compared to nisin and up to 2500-1700 IU mL-1 as compared to fungicidal antibiotic nistatin. In our experiments we used raw smoked sausages that were infected with fungi. The identification of this infection showed the presence of Eurotium repens de Bary on the sausages. Treatment of the raw smoked sausages with cultural broth of L.lactis ssp. lactis 194 and K-205 inhibited growth of these microorganisms. After treatment the sausages had longer shelf-life and was in accordance with basal production data (Russian State Standard Specification 16131-86. Conclusion: The results of this study indicated that the treatment with

  5. Lactococcus lactis productor de bacteriocina utilizable como cultivo iniciador para acelerar la maduración de queso

    OpenAIRE

    Martínez-Cuesta, M. Carmen; Requena, Teresa; Peláez, Carmen

    2003-01-01

    Lactococcus lactis productor de bacteriocina utilizable como cultivo iniciador para acelerar la maduración de queso. La presente invención describe un procedimiento de producción de transconjugantes de Lactococcus lactis (se describe en detalle el Lactococcus lactis CECT5367) productores de bacteriocina y por tanto inmune a ella. Este microorganismo y otros similares pueden utilizarse como cultivos iniciadores para la elaboración de queso semiduro produciendo quesos de buena calidad organolép...

  6. Application of the ligase chain reaction to the detection of nisinA and nisinZ genes in Lactococcus lactis ssp. lactis.

    Science.gov (United States)

    Ward, L J; Brown, J C; Davey, G P

    1994-03-15

    This paper reports on the application of the ligase chain reaction (LCR) to the specific detection of variants of the nisin structural gene (nisinA and nisinZ) in nisin producing strains of Lactococcus lactis ssp lactis. The LCR assay was used to screen nisin producing strains to determine which form of the nisin structural gene they contained. This method of differentiating the nisin structural gene variants provides a useful alternative to the only other available genetic differentiation, that of sequencing the gene.

  7. Influence of Stress Treatments on the Resistance of Lactococcus lactis to Freezing and Freeze-Drying

    OpenAIRE

    Lin, Chan

    1998-01-01

    This study investigated the effect of cold, heat, or osmotic shock treatment on the resistance of L. lactis subsp. cremoris MM160 and MM310 and Lactococcus lactis subsp. lactis MM210 and FG2 cheese starter bacteria to freezing and freeze-drying. The ability to withstand freezing at -60°C for 24 h was variable among lactococci, but resistance to this treatment was significantly improved (P < 0.05) in most strains by a 2-h cold shock at l0°C or a 25-min heat shock at 39°C (L. lactis subsp. crem...

  8. Antimicrobial activity of Lactobacillus and Bifidobacterium strains against pathogenic microorganisms “in vitro”Atividade antimicrobiana de Lactobacillus e Bifodobacterium frente a microrganismos patogênicos “in vitro”

    Directory of Open Access Journals (Sweden)

    Giselle Nobre Costa

    2012-10-01

    Full Text Available Lactobacilli and bifidobacteria have a long history of safe use in foods. These bacteria have biotechnological characteristics of interest such as the inhibition of pathogens. In this work, two lactobacilli strain and a bifidobacterium strain isolated from human gut were evaluated concerning to their ability to inhibit pathogenic microorganisms in foods by diffusion agar tests. Moreover, we assessed the metabolites produced in culture broth under static and shaking growth to simulate anaerobiosis and aerobiosis conditions, respectively. L. acidophilus LA5, L. plantarum DCTA 8420 and B. lactis DCTA 8724 showed ability to inhibit S. aureus FRI 196, strains producer toxins A and D, as well as B. cereus ATCC 25923, E. coli ATCC 25922 and S. Enteritidis, whose inhibition halos reached, on average, 24 mm in diameter. In the agar diffusion method with concentrated culture medium, it was possible to observe the effect of oxygen on the production of toxic substances. This result showed that cultivation of Lactobacillus under aerobic conditions seems to exert greater inhibitory effect, whereas for Bifidobacterium strain the effect was the opposite.Lactobacilos e bifidobactérias apresentam um longo histórico de uso seguro em alimentos, além de apresentarem características de interesse biotecnológico como a inibição de patógenos. Neste trabalho duas linhagens de lactobacilos e uma de bifidobactéria, isoladas do intestino humano, foram avaliadas em testes de difusão em ágar, quanto à capacidade de inibição de microrganismos patogênicos de ocorrência comuns em toxinfecções alimentares. Adicionalmente, foram avaliados os metabólitos produzidos em caldo de cultivo estático e em agitação para simular condições de anaerobiose a aerobiose, respectivamente. As três bactérias, L. acidophilus LA5, L. plantarum DCTA 8420 e B. lactis DCTA 8724 apresentaram capacidade de inibição para S. aureus FRI 196 linhagem produtora de toxinas A e D

  9. Bifidobacterium moukalabense sp. nov., isolated from the faeces of wild west lowland gorilla (Gorilla gorilla gorilla).

    Science.gov (United States)

    Tsuchida, Sayaka; Takahashi, Shunsuke; Nguema, Pierre Philippe Mbehang; Fujita, Shiho; Kitahara, Maki; Yamagiwa, Juichi; Ngomanda, Alfred; Ohkuma, Moriya; Ushida, Kazunari

    2014-02-01

    Gram-staining-positive anaerobic rods were isolated from the faeces of a wild lowland gorilla (Gorilla gorilla gorilla) in Moukalaba-Doudou National Park, Gabon, and strain GG01(T) was taxonomically investigated. Based on phylogenetic analyses and specific phenotypic characteristics, the strain belonged to the genus Bifidobacterium. Phylogenetic analysis of its 16S rRNA gene sequence revealed that strain GG01(T) formed a single monophyletic cluster and had a distinct line of descent. Based on 16S rRNA gene sequence similarity, the type strains of Bifidobacterium catenulatum JCM 1194(T) (98.3%) and Bifidobacterium pseudocatenulatum (98.1%) JCM 1200(T) were the most closely related to this novel strain, although it was clear that they belonged to different species. hsp60 sequences also supported these relationships. The DNA G+C content of this novel strain was 60.1 mol%. Bifidobacterium moukalabense sp. nov. (type strain GG01(T) = JCM 18751(T) = DSM 27321(T)) is proposed.

  10. Imbalances in faecal and duodenal Bifidobacterium species composition in active and non-active coeliac disease

    Directory of Open Access Journals (Sweden)

    Sanz Yolanda

    2008-12-01

    Full Text Available Abstract Background Gut bifidobacteria are believed to influence immune-related diseases. The objective of this study was to assess the possible relationships between the gut bifidobacteria composition and coeliac disease (CD in children. A total of 48 faecal samples (30 and 18 samples from active and no active CD patients, respectively and 33 duodenal biopsy specimens of CD patients (25 and 8 samples from active and non-active CD patients, respectively were analysed. Samples (30 faecal samples and 8 biopsies from a control age-matched group of children were also included for comparative purposes. Gut Bifidobacterium genus and species were analyzed by real-time PCR. Results Active and non-active CD patients showed lower numbers of total Bifidobacterium and B. longum species in faeces and duodenal biopsies than controls, and these differences were particularly remarkable between active CD patients and controls. B. catenulatum prevalence was higher in biopsies of controls than in those of active and non-active CD patients, whereas B. dentium prevalence was higher in faeces of non-active CD patients than in controls. Correlations between levels of Bifidobacterium and B. longum species in faecal and biopsy samples were detected in both CD patients and controls. Conclusion Reductions in total Bifidobacterium and B. longum populations were associated with both active and non-active CD when compared to controls. These bacterial groups could constitute novel targets for adjuvant dietary therapies although the confirmation of this hypothesis would require further investigations.

  11. Influence of bifidobacterium quadruple preparations on immunity function and cytokine of neonatal hyperbilirubinemia

    Institute of Scientific and Technical Information of China (English)

    Juan Zhang; Xiao-Qin Liang

    2016-01-01

    Objective:To investigate the influence of bifidobacterium quadruple preparations on immunity function and cytokine of neonatal hyperbilirubinemia.Methods:A total of 92 neonates with hyperbilirubinemia were randomly divided into observation group and control group with 46 cases of each group. The control group received conventional therapy, and the observation group was treated with bifidobacterium quadruple preparations on the basis of the control group. The influence of bifidobacterium quadruple preparations on immunity function and cytokine of the two groups were compared.Results:There was no significantly difference in Bilirubin before the treatment between the two groups (P>0.05), but the observation group was significantly lower than the control group 3, 5, 7 d after the treatment (P<0.01); CD3, CD4, CD4/CD8, IL-6, IL-8 of the observation group were higher than the control group after the treatment (P<0.01), but CD8 was lower than the control group (P<0.01).Conclusions:Bifidobacterium quadruple preparations can improve the immune function, and reduce serum bilirubin levels in neonates with hyperbilirubinemia.

  12. Bifidobacterium bifidum Actively Changes the Gene Expression Profile Induced by Lactobacillus acidophilus in Murine Dendritic Cells

    DEFF Research Database (Denmark)

    Weiss, Gudrun Margarethe; Rasmussen, Simon; Fink, Lisbeth Nielsen

    2010-01-01

    cytokine IL-12 in DC, whereas bifidobacteria do not induce IL-12 but inhibit the IL-12 production induced by lactobacilli. In the present study, genome-wide microarrays were used to investigate the gene expression pattern of murine DC stimulated with Lactobacillus acidophilus NCFM and Bifidobacterium...

  13. Structural rearrangements of sucrose phosphorylase from Bifidobacterium adolescentis during sucrose conversion

    DEFF Research Database (Denmark)

    Mirza, Osman; Henriksen, Lars Skov; Sprogøe, Desiree

    2006-01-01

    The reaction mechanism of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) was studied by site-directed mutagenesis and x-ray crystallography. An inactive mutant of BiSP (E232Q) was co-crystallized with sucrose. The structure revealed a substrate-binding mode comparable with that seen...

  14. Specific detection and analysis of a probiotic Bifidobacterium strain in infant feces

    NARCIS (Netherlands)

    Kok, RG; DeWaal, A; Schut, F; Welling, GW; Weenk, G; Hellingwerf, KJ

    1996-01-01

    For specific detection of the probiotic Bifidobacterium sp. strain LW420 in infant feces and for rapid quality control of this strain in culture, three strain-specific 16S rRNA gene-targeted primers have been developed. These primers allow specific detection of the organism via PCR. Specificity of t

  15. Discovering novel bile protection systems in Bifidobacterium breve UCC2003 through functional genomics.

    NARCIS (Netherlands)

    Ruiz, L.; Zomer, A.L.; O'Connell-Motherway, M.; Sinderen, D. van; Margolles, A.

    2012-01-01

    Tolerance of gut commensals to bile salt exposure is an important feature for their survival in and colonization of the intestinal environment. A transcriptomic approach was employed to study the response of Bifidobacterium breve UCC2003 to bile, allowing the identification of a number of bile-induc

  16. Susceptibility of human and probiotic Bifidobacterium spp. to selected antibiotics as determined by the Etest method

    NARCIS (Netherlands)

    Matto, J.; Hoek, van A.H.A.M.; Domig, K.J.; Saarela, M.; Flórez, A.B.; Brockmann, E.; Amtmann, E.; Mayo, B.; Aarts, H.J.M.; Danielsen, M.

    2007-01-01

    This study reports the antibiotic susceptibility of 203 strains representing human or probiotic associated Bifidobacterium species as determined by the Etest method. Strains showing minimum inhibitory concentration (MIC) for tetracycline >= 16 mu g mL(-1) were detected in all studied Bifidobacter

  17. Applications of Microencapsulated Bifidobacterium Longum with Eleutherine Americana in Fresh Milk Tofu and Pineapple Juice

    OpenAIRE

    Atchara N Phoem; Suphitchaya Chanthachum; Supayang P Voravuthikunchai

    2015-01-01

    Bifidobacterium longum was microencapsulated by extrusion technique and added in fresh milk tofu and pineapple juice. Microencapsulation of B. longum with Eleutherine americana extract, oligosaccharides extract, and commercial fructo-oligosaccharides was assessed for the bacterial survival after sequential exposure to simulated gastric and intestinal juices, and refrigeration storage. Microencapsulated B. longum with the extract and oligosaccharides extract in the food products showed better ...

  18. Bifidobacterium breve - HT-29 cell line interaction: modulation of TNF-a induced gene expression

    NARCIS (Netherlands)

    Boesten, R.J.; Vos, de W.M.; Schuren, F.H.J.; Willemsen, L.E.M.; Knol, J.

    2011-01-01

    To provide insight in the molecular basis for intestinal host-microbe interactions, we determined the genome-wide transcriptional response of human intestinal epithelial cells following exposure to cells of Bifidobacterium breve. To select an appropriate test system reflecting inflammatory condition

  19. Bifidobacterium breve-HT-29 cell line interaction: Modulation of TNF-a induced gene expression

    NARCIS (Netherlands)

    Boesten, R.J.; Schuren, F.H.J.; Willemsen, L.E.M.; Vriesema, A.; Knol, J.; Vos, W.M. de

    2011-01-01

    To provide insight in the molecular basis for intestinal host-microbe interactions, we determined the genome-wide transcriptional response of human intestinal epithelial cells following exposure to cells of Bifidobacterium breve. To select an appropriate test system reflecting inflammatory condition

  20. Physico-chemical and transglucosylation properties of recombinant sucrose phosphorylase from Bifidobacterium adolescentis DSM20083.

    NARCIS (Netherlands)

    Broek, van den L.A.M.; Boxtel, van E.L.; Kievit, R.P.; Verhoef, R.P.; Beldman, G.; Voragen, A.G.J.

    2004-01-01

    Clones of a genomic library of Bifidobacterium adolescentis were grown in minimal medium with sucrose as sole carbon source. An enzymatic fructose dehydrogenase assay was used to identify sucrose-degrading enzymes. Plasmids were isolated from the positive colonies and sequence analysis revealed that

  1. Rapid detection of human fecal contamination in estuarine environments by PCR targeting of Bifidobacterium adolescentis

    NARCIS (Netherlands)

    King, Eric L.; Bachoon, Dave S.; Gates, Keith W.

    2007-01-01

    Detection of Bifidobacterium adolescentis was used as an effective genetic marker of human fecal contamination in Georgia estuaries. Enterococci enumerations on mEI media indicated that a tributary to the Little Satilla River with 516 CFU/100 ml was the most polluted of all the rivers tested. Extrac

  2. Occurrence and Diversity of CRISPR-Cas Systems in the Genus Bifidobacterium.

    Directory of Open Access Journals (Sweden)

    Alexandra E Briner

    Full Text Available CRISPR-Cas systems constitute adaptive immune systems for antiviral defense in bacteria. We investigated the occurrence and diversity of CRISPR-Cas systems in 48 Bifidobacterium genomes to gain insights into the diversity and co-evolution of CRISPR-Cas systems within the genus and investigate CRISPR spacer content. We identified the elements necessary for the successful targeting and inference of foreign DNA in select Type II CRISPR-Cas systems, including the tracrRNA and target PAM sequence. Bifidobacterium species have a very high frequency of CRISPR-Cas occurrence (77%, 37 of 48. We found that many Bifidobacterium species have unusually large and diverse CRISPR-Cas systems that contain spacer sequences showing homology to foreign genetic elements like prophages. A large number of CRISPR spacers in bifidobacteria show perfect homology to prophage sequences harbored in the chromosomes of other species of Bifidobacterium, including some spacers that self-target the chromosome. A correlation was observed between strains that lacked CRISPR-Cas systems and the number of times prophages in that chromosome were targeted by other CRISPR spacers. The presence of prophage-targeting CRISPR spacers and prophage content may shed light on evolutionary processes and strain divergence. Finally, elements of Type II CRISPR-Cas systems, including the tracrRNA and crRNAs, set the stage for the development of genome editing and genetic engineering tools.

  3. Diversity analysis of dairy and nondairy Lactococcus lactis isolates, using a novel multilocus sequence analysis scheme and (GTG)5-PCR fingerprinting

    NARCIS (Netherlands)

    Rademaker, J.L.W.; Herbet, H.; Starrenburg, M.J.C.; Naser, S.M.; Gevers, D.; Kelly, W.J.; Hugenholtz, J.; Swings, J.; van Hylckama Vlieg, J.E.T.

    2007-01-01

    The diversity of a collection of 102 lactococcus isolates including 91 Lactococcus lactis isolates of dairy and nondairy origin was explored using partial small subunit rRNA gene sequence analysis and limited phenotypic analyses. A subset of 89 strains of L. lactis subsp. cremoris and L. lactis subs

  4. Tripeptidase Gene (pepT) of Lactococcus lactis : Molecular Cloning and Nucleotide Sequencing of pepT and Construction of a Chromosomal Deletion Mutant

    NARCIS (Netherlands)

    Mierau, Igor; Haandrikman, Alfred J.; Velterop, Odilia; Tan, Paris S.T.; Leenhouts, Kees L.; Konings, Wilhelmus; Venema, Gerhardus; Kok, Jan

    1994-01-01

    The gene encoding a tripeptidase (pepT) of lactococcus lactis subsp. cremoris (formerly subsp. lactis) MG1363 was cloned from a genomic library in pUC19 and subsequently sequenced. The tripeptidase of L. lactis was shown to be homologous to PepT of Salmonella typhimurium with 47.4% identity in the d

  5. The prophylactic effect of probiotic Enterococcus lactis IW5 against different human cancer cells

    Directory of Open Access Journals (Sweden)

    YOUSEF eNAMI

    2015-11-01

    Full Text Available Enterococcus lactis IW5 was obtained from human gut and the potential probiotic characteristics of this organism were then evaluated. Results showed that this strain was highly resistant to low pH and high bile salt and adhered strongly to Caco-2 human epithelial colorectal cell lines. The supernatant of E. lactis IW5 strongly inhibited the growth of several pathogenic bacteria and decreased the viability of different cancer cells, such as HeLa, AGS, HT-29, and MCF-7. Conversely, E. lactis IW5 did not inhibit the viability of normal FHs-74 cells. This strain did not generate toxic enzymes, including β-glucosidase, β-glucuronidase, and N-acetyl-β-glucosaminidase and was highly susceptible to ampicillin, gentamycin, penicillin, vancomycin, clindamycin, sulfamethoxazol, and chloramphenicol but resistant to erythromycin and tetracyclin. This study provided evidence for the effect of E. lactis IW5 on cancer cells. Therefore, E. lactis IW5, as a bioactive therapeutics, should be subjected to other relevant tests to verify the therapeutic suitability of this strain for clinical applications.

  6. Bacteriocins produced by wild Lactococcus lactis strains isolated from traditional, starter-free cheeses made of raw milk.

    Science.gov (United States)

    Alegría, Angel; Delgado, Susana; Roces, Clara; López, Belén; Mayo, Baltasar

    2010-09-30

    Sixty bacterial strains were encountered by random amplification of polymorphic DNA (RAPD) and repetitive extragenic palindromic (REP) typing in a series of 306 Lactococcus lactis isolates collected during the manufacturing and ripening stages of five traditional, starter-free cheeses made from raw milk. Among the 60 strains, 17 were shown to produce bacteriocin-like compounds in both solid and liquid media. At a genotypic level, 16 of the strains were identified by molecular methods as belonging to L. lactis subsp. lactis and one to L. lactis subsp. cremoris. Among the L. lactis subsp. lactis strains, phenotypic and genetic data determined that eleven produced either nisin A (nine strains) or nisin Z (two strains), and that five produced lactococcin 972. Variable levels of the two bacteriocins were produced by different strains. In addition, nisin was shown to be produced in inexpensive, dairy- and meat-based media, which will allow the practical application of its producing strains in industrial processes. Specific PCR and nucleotide and deduced amino acid sequence analysis identified the inhibitor produced by the single L. lactis subsp. cremoris isolate as a lactococcin G-like bacteriocin. Beyond the use of bacteriocins as functional ingredients for the biopreservation of foods, the newly identified bacteriocin-producing L. lactis strains from traditional cheeses may also be useful for designing starter cultures with protective properties and/or adjunct cultures for accelerating cheese ripening.

  7. Fermentation-induced variation in heat and oxidative stress phenotypes of Lactococcus lactis MG1363 reveals transcriptome signatures for robustness

    NARCIS (Netherlands)

    A.R. Dijkstra; W. Alkema; M.J.C. Starrenburg; J. Hugenholtz; S.A.F.T. van Hijum; P.A. Bron

    2014-01-01

    Background: Lactococcus lactis is industrially employed to manufacture various fermented dairy products. The most cost-effective method for the preservation of L. lactis starter cultures is spray drying, but during this process cultures encounter heat and oxidative stress, typically resulting in low

  8. Evolution of Lactococcus lactis phages within a cheese factory.

    Science.gov (United States)

    Rousseau, Geneviève M; Moineau, Sylvain

    2009-08-01

    We have sequenced the double-stranded DNA genomes of six lactococcal phages (SL4, CB13, CB14, CB19, CB20, and GR7) from the 936 group that were isolated over a 9-year period from whey samples obtained from a Canadian cheese factory. These six phages infected the same two industrial Lactococcus lactis strains out of 30 tested. The CB14 and GR7 genomes were found to be 100% identical even though they were isolated 14 months apart, indicating that a phage can survive in a cheese plant for more than a year. The other four genomes were related but notably different. The length of the genomes varied from 28,144 to 32,182 bp, and they coded for 51 to 55 open reading frames. All five genomes possessed a 3' overhang cos site that was 11 nucleotides long. Several structural proteins were also identified by nano-high-performance liquid chromatography-tandem mass spectrometry, confirming bioinformatic analyses. Comparative analyses suggested that the most recently isolated phages (CB19 and CB20) were derived, in part, from older phage isolates (CB13 and CB14/GR7). The organization of the five distinct genomes was similar to the previously sequenced lactococcal phage genomes of the 936 group, and from these sequences, a core genome was determined for lactococcal phages of the 936 group.

  9. Geraniol dehydrogenase, the key enzyme in biosynthesis of the alarm pheromone, from the astigmatid mite Carpoglyphus lactis (Acari: Carpoglyphidae).

    Science.gov (United States)

    Noge, Koji; Kato, Makiko; Mori, Naoki; Kataoka, Michihiko; Tanaka, Chihiro; Yamasue, Yuji; Nishida, Ritsuo; Kuwahara, Yasumasa

    2008-06-01

    Geraniol dehydrogenase (GeDH), which plays an important role in the biosynthesis of neral, an alarm pheromone, was purified from the astigmatid mite Carpoglyphus lactis. The enzyme was obtained in an apparently homogeneous and active form after 1879-fold purification through seven steps of chromatography. Car. lactis GeDH was determined to be a monomer in its active form with a relative molecular mass of 42 800, which is a unique subunit structure in comparison with already established alcohol dehydrogenases. Car. lactis GeDH oxidized geraniol into geranial in the presence of NAD+. NADP+ was ineffective as a cofactor, suggesting that Car. lactis GeDH is an NAD+-dependent alcohol dehydrogenase. The optimal pH and temperature for geraniol oxidation were determined to be pH 9.0 and 25 degrees C, respectively. The Km values for geraniol and NAD+ were 51.0 microm and 59.5 microm, respectively. Car. lactis GeDH was shown to selectively oxidize geraniol, whereas its geometrical isomer, nerol, was inert as a substrate. The high specificity for geraniol suggests that Car. lactis GeDH specializes in the alarm pheromone biosynthesis of Car. lactis. Car. lactis GeDH is composed of 378 amino acids. Structurally, Car. lactis GeDH showed homology with zinc-dependent alcohol dehydrogenases found in mammals and a mosquito (36.6-37.6% identical), and the enzyme was considered to be a member of the medium-chain dehydrogenase/reductase family, in view of the highly conserved sequences of zinc-binding and NAD+-binding sites. Phylogenetic analyses indicate that Car. lactis GeDH could be categorized as a new class, different from other established alcohol dehydrogenases.

  10. An exoproteome approach to monitor safety of a cheese-isolated Lactococcus lactis

    DEFF Research Database (Denmark)

    Genovese, Federica; Coïsson, Jean Daniel; Majumder, Avishek

    2013-01-01

    The safety of the cheese-isolated and potential starter Lactococcus lactis 11D was explored by means of an extracellular proteomic study. A preliminary analysis showed good caseification/proteolytic behavior of the strain, absence of production of biogenic amines and good survival at acidic p...... isomerase were abundant in the L. lactis 11D exoproteome. These proteins play a role in bacterial aggregation and in bacteria–fungi interactions, therefore their presence may indicate a good competition potential of the strain against other microorganisms in both food and the gastrointestinal habitat....... This is to our knowledge the first extracellular proteomic mapping of L. lactis with relevance for bacterial strain-typing in food safety....

  11. Physiochemical parameters optimization for enhanced nisin production by Lactococcus lactis (MTCC 440

    Directory of Open Access Journals (Sweden)

    Puspadhwaja Mall

    2010-02-01

    Full Text Available The influence of various physiochemical parameters on the growth of Lactococcus lactis sub sp. lactis MTCC 440 was studied at shake flask level for 20 h. Media optimization (MRS broth was studied to achieve enhanced growth of the organism and also nisin production. Bioassay of nisin was done with agar diffusion method using Streptococcus agalactae NCIM 2401 as indicator strain. MRS broth (6%, w/v with 0.15μg/ml of nisin supplemented with 0.5% (v/v skimmed milk was found to be the best for nisin production as well as for growth of L lactis. The production of nisin was strongly influenced by the presence of skimmed milk and nisin in MRS broth. The production of nisin was affected by the physical parameters and maximum nisin production was at 30(0C while the optimal temperature for biomass production was 37(0C.

  12. Bifidobacterium aesculapii sp. nov., from the faeces of the baby common marmoset (Callithrix jacchus).

    Science.gov (United States)

    Modesto, M; Michelini, S; Stefanini, I; Ferrara, A; Tacconi, S; Biavati, B; Mattarelli, P

    2014-08-01

    Six Gram-positive-staining, microaerophilic, non-spore-forming, fructose-6-phosphate phosphoketolase-positive bacterial strains with a peculiar morphology were isolated from faecal samples of baby common marmosets (Callithrix jacchus). Cells of these strains showed a morphology not reported previously for a bifidobacterial species, which resembled a coiled snake, always coiled or ring shaped or forming a 'Y' shape. Strains MRM 3/1(T) and MRM 4/2 were chosen as representative strains and characterized further. The bacteria utilized a wide range of carbohydrates and produced urease. Glucose was fermented to acetate and lactate. Strain MRM 3/1(T) showed a peptidoglycan type unique among members of the genus Bifidobacterium. The DNA base composition was 64.7 mol% G+C. Almost-complete 16S rRNA, hsp60, clpC and rpoB gene sequences were obtained and phylogenetic relationships were determined. Comparative analysis of 16S rRNA gene sequences showed that strains MRM 3/1(T) and MRM 4/2 had the highest similarities to Bifidobacterium scardovii DSM 13734(T) (94.6%) and Bifidobacterium stellenboschense DSM 23968(T) (94.5%). Analysis of hsp60 showed that both strains were closely related to B. stellenboschense DSM 23968(T) (97.5% similarity); however, despite this high degree of similarity, our isolates could be distinguished from B. stellenboschense DSM 23968(T) by low levels of DNA-DNA relatedness (30.4% with MRM 3/1(T)). Strains MRM 3/1(T) and MRM 4/2 were located in an actinobacterial cluster and were more closely related to the genus Bifidobacterium than to other genera in the family Bifidobacteriaceae. On the basis of these results, strains MRM 3/1(T) and MRM 4/2 represent a novel species within the genus Bifidobacterium, for which the name Bifidobacterium aesculapii sp. nov. is proposed; the type strain is MRM 3/1(T) ( = DSM 26737(T) = JCM 18761(T)).

  13. Physiological function of the maltose operon regulator, MalR, in Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    Rådström Peter

    2002-09-01

    Full Text Available Abstract Background Maltose metabolism is initiated by an ATP-dependent permease system in Lactococcus lactis. The subsequent degradation of intracellular maltose is performed by the concerted action of Pi-dependent maltose phosphorylase and β-phosphoglucomutase. In some Gram-positive bacteria, maltose metabolism is regulated by a maltose operon regulator (MalR, belonging to the LacI-GalR family of transcriptional regulators. A gene presumed to encode MalR has been found directly downstream the maltose phosphorylase-encoding gene, malP in L. lactis. The purpose of this study was to investigate the physiological role of the MalR protein in maltose metabolism in L. lactis. Results A L. lactis ssp. lactis mutant, TMB5004, deficient in the putative MalR protein, was physiologically characterised. The mutant was not able to ferment maltose, while its capability to grow on glucose as well as trehalose was not affected. The activity of maltose phosphorylase and β-phosphoglucomutase was not affected in the mutant. However, the specific maltose uptake rate in the wild type was, at its lowest, five times higher than in the mutant. This difference in maltose uptake increased as the maltose concentration in the assay was increased. Conclusion According to amino acid sequence similarities, the presumed MalR is a member of the LacI-GalR family of transcriptional regulators. Due to the suggested activating effect on maltose transport and absence of effect on the activities of maltose phosphorylase and β-phosphoglucomutase, MalR of L. lactis is considered rather as an activator than a repressor.

  14. Bioelectrochemical Mn(II) leaching from manganese ore by Lactococcus lactis SK071115.

    Science.gov (United States)

    Jeon, Bo Young; Park, Doo Hyun

    2011-02-01

    L. lactis sk071115 has been shown to grow more actively and generate lower levels of lactate in glucose-defined medium with nitrate than in medium with Mn(IV). By adding Mn(IV) to a L. lactis culture, lactate production was relatively reduced in combination with Mn(II) production, but cell mass production levels did not increase. Both cell-free extract and intact L. lactis cells reacted electrochemically with Mn(IV) but did not react with Mn(II) upon cyclic voltammetry using neutral red (NR) as an electron mediator. A modified graphite felt cathode with NR (NR-cathode) was employed to induce electrochemical reducing equivalence for bacterial metabolism. Cell-free L. lactis extract catalyzed the reduction of Mn(IV) to Mn(II) under both control and electrochemical reduction conditions; however, the levels of Mn(II) generated under electrochemical reduction conditions were approximately 4 times those generated under control conditions. The levels of Mn(II) generated by the catalysis of L. lactis immobilized in the NR-cathode (L-NR-cathode) under electrochemical reduction conditions were more than 4 times that generated under control conditions. Mn(II) production levels were increased by approximately 2.5 and 4.5 times by the addition of citrate to the reactant under control and electrochemical reduction conditions, respectively. The cumulative Mn(II) produced from manganese ore by catalysis of the L-NR-cathode for 30 days reached levels of approximately 3,800 and 16,000 mg/l under control and electrochemical reduction conditions, respectively. In conclusion, the electrochemical reduction reaction generated by the NR-cathode activated the biochemical reduction of Mn(IV) to Mn(II) by L. lactis.

  15. Secretion of biologically active interferon-gamma inducible protein-10 (IP-10 by Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    Saucedo-Cardenas Odila

    2008-07-01

    Full Text Available Abstract Background Chemokines are a large group of chemotactic cytokines that regulate and direct migration of leukocytes, activate inflammatory responses, and are involved in many other functions including regulation of tumor development. Interferon-gamma inducible-protein-10 (IP-10 is a member of the C-X-C subfamily of the chemokine family of cytokines. IP-10 specifically chemoattracts activated T lymphocytes, monocytes, and NK cells. IP-10 has been described also as a modulator of other antitumor cytokines. These properties make IP-10 a novel therapeutic molecule for the treatment of chronic and infectious diseases. Currently there are no suitable live biological systems to produce and secrete IP-10. Lactococcus lactis has been well-characterized over the years as a safe microorganism to produce heterologous proteins and to be used as a safe, live vaccine to deliver antigens and cytokines of interest. Here we report a recombinant strain of L. lactis genetically modified to produce and secrete biologically active IP-10. Results The IP-10 coding region was isolated from human cDNA and cloned into an L. lactis expression plasmid under the regulation of the pNis promoter. By fusion to the usp45 secretion signal, IP-10 was addressed out of the cell. Western blot analysis demonstrated that recombinant strains of L. lactis secrete IP-10 into the culture medium. Neither degradation nor incomplete forms of IP-10 were detected in the cell or supernatant fractions of L. lactis. In addition, we demonstrated that the NICE (nisin-controlled gene expression system was able to express IP-10 "de novo" even two hours after nisin removal. This human IP-10 protein secreted by L. lactis was biological active as demonstrated by Chemotaxis assay over human CD3+T lymphocytes. Conclusion Expression and secretion of mature IP-10 was efficiently achieved by L. lactis forming an effective system to produce IP-10. This recombinant IP-10 is biologically active as

  16. Oxidative Stress at High Temperatures in Lactococcus lactis Due to an Insufficient Supply of Riboflavin

    DEFF Research Database (Denmark)

    Shen, Jing; Solem, Christian; Jensen, Peter Ruhdal;

    2013-01-01

    . These results indicate that L. lactis suffers from heat-induced oxidative stress at increased temperatures. A decrease in intracellular flavin adenine dinucleotide (FAD), which is derived from riboflavin, was observed with increasing growth temperature, but the presence of riboflavin made the decrease smaller...... riboflavin to the medium, it was possible to improve growth and oxygen consumption at 37°C, and this also normalized the [ATP]-to-[ADP] ratio. A codon-optimized redox-sensitive green fluorescent protein (GFP) was introduced into L. lactis and revealed a more oxidized cytoplasm at 37°C than at 30°C...

  17. Molecular and phenotypic traits of in-vitro-selected mutants of Bifidobacterium resistant to rifaximin.

    Science.gov (United States)

    Vitali, Beatrice; Turroni, Silvia; Serina, Stefania; Sosio, Margherita; Vannini, Lucia; Candela, Marco; Guerzoni, Maria Elisabetta; Brigidi, Patrizia

    2008-06-01

    Nucleotide mutations inside a core region of the rpoB gene, encoding the beta subunit of RNA polymerase, were found in rifaximin-resistant mutants of Bifidobacterium. Five different missense mutations of codons 513, 516, 522 and 529 were identified. Further aspects of rifaximin resistance were investigated, using Bifidobacterium infantis BI07 as a model strain. Partial resistance of RNA polymerase of a BI07 mutant at a rifaximin concentration >10 microg/mL was observed by cell-free transcription assay. Mass spectrometry detection of rifaximin in the cellular pellet of the BI07 resistant mutant, as well as changes in biosynthesis of saturated and cyclopropane fatty acids during growth, suggested a reduction in membrane permeability for the antibiotic moiety.

  18. Microencapsulation and Fermentation of Lactobacillus acidophilus LA-5 and Bifidobacterium BB-12

    Directory of Open Access Journals (Sweden)

    Maryam Yari

    2015-09-01

    Full Text Available Because of poor survival of probiotic bacteria, microencapsulation evolved from the immobilized cell culture technology used in the biotechnological industry. Two probiotic strains, Bifidobacterium (BB-12 and Lactobacillus acidophilus (LA-5 were immobilized in calcium alginate by extrusion method. Encapsulation parameters and efficacy of this method were evaluated. Growth factors of these two bacteria were also measured by culturing in 10-L fermenter. Growth curves were obtained with respect to optical density and dry biomass weight. Encapsulation yield was over than 60% in each experiment. Scanning electron microscopy (SEM of Entrapment of cells in alginate matrix and cross-sections of dried bead were obtained and illustrated. Bifidobacterium have been shown better biotechnological properties.

  19. Optimizing the Chemical Compositions of Protective Agents for Freeze-drying Bifidobacterium longum BIOMA 5920

    Institute of Scientific and Technical Information of China (English)

    杨婵媛; 朱晓丽; 范代娣; 米钰; 骆艳娥; 惠俊峰; 苏然

    2012-01-01

    Freeze drying has a deleterious effect on the viability of microorganisms. In front of this difficulty, the present study adopts response surface methodology to optimize the chemical compositions of protective agents to seek for maximum viability of Bifidobacterium longum BIOMA 5920 during freeze-drying. Through the compara- tive analysis of single protectant, the complex protective agents show better effect on the Bifidobacterium viability. Human-like collagen (HLC), trehalose and glycerol are confirmed as significant factors by Box-Behnken Design. The optimized formula for these three variables is tested as follows: HLC 1.23%, trehalose 11.50% and glycerol 4.65%. Under this formula, the viability is 88.23%, 39.67% higher in comparison to the control. The viable count is 1.07×10 9 cfu·g-1 , greatly exceeding the minimum viable count requirement (10 6 cfu·g-1 ).

  20. POSSIBLE ROLE OF PROBIOTIC MICROORGANISMS OF BIFIDOBACTERIUM AND LACTOBACILLUS GENUS IN PATHOGENESIS OF AUTOIMMUNE THYROID DISEASES

    Directory of Open Access Journals (Sweden)

    E. P. Kiseleva

    2010-01-01

    Full Text Available It was revealed, that, in blood samples of the patients with autoimmune thyroid diseases, serum antibodies against cell-free fraction of Bifidobacterium bifidum 791 and Lactobacillus plantarum B-01 were detected, respectively, in 71 and 63% of cases, that being two-fold higher than appropriate frequencies in healthy blood donors. An evidence was obtained that presence of some components specifically reacting with autoantibodies against thyroid peroxidase and thyroglobulin on the surface of the microorganisms cells and competing for binding of these immunoglobulins with thyroid antigens. One may also suggest a presence of bacterial components, interacting with thyroid peroxidase. The data obtained let us suggest that probiotic microorganisms of Bifidobacterium and Lactobacillus genus could take part in pathogenesis of autoimmune thyroid diseases, by means of molecular mimicry mechanisms.

  1. Efficacy of oral Bifidobacterium bifidum ATCC 29521 on microflora and antioxidant in mice.

    Science.gov (United States)

    Wang, Bao-gui; Xu, Hai-bo; Xu, Feng; Zeng, Zhe-ling; Wei, Hua

    2016-03-01

    This study aimed to examine whether Bifidobacterium bifidum ATCC 29521, a species of colonic microflora in humans, is involved in the intestinal tract of mice. This study was also conducted to determine the antioxidant activity of this species by evaluating different microbial populations and reactive oxygen species isolated from feces and intestinal contents for 28 days of oral administration. Microbial diversities were assessed through bacterial culture techniques, PCR-DGGE, and real-time PCR. This study showed that the intake of B. bifidum ATCC 29521 significantly (p stress were also determined. Results indicated that B. bifidum ATCC 29521 elicits a beneficial effect on murine gut microbiota and antioxidant activities compared with the control samples. This species can be considered as a potential bioresource antioxidant to promote health. Bifidobacterium bifidum ATCC 29521 may also be used as a promising material in microbiological and food applications.

  2. In Vitro Kinetic Analysis of Oligofructose Consumption by Bacteroides and Bifidobacterium spp. Indicates Different Degradation Mechanisms

    OpenAIRE

    Van der Meulen, Roel; Makras, Lefteris; Verbrugghe, Kristof; Adriany, Tom; De Vuyst, Luc

    2006-01-01

    The growth of pure cultures of Bacteroides thetaiotaomicron LMG 11262 and Bacteroides fragilis LMG 10263 on fructose and oligofructose was examined and compared to that of Bifidobacterium longum BB536 through in vitro laboratory fermentations. Gas chromatography (GC) analysis was used to determine the different fractions of oligofructose and their degradation during the fermentation process. Both B. thetaiotaomicron LMG 11262 and B. fragilis LMG 10263 were able to grow on oligofructose as fas...

  3. Probiotic Bifidobacterium longum alters gut luminal metabolism through modification of the gut microbial community

    OpenAIRE

    Hirosuke Sugahara; Toshitaka Odamaki; Shinji Fukuda; Tamotsu Kato; Jin-zhong Xiao; Fumiaki Abe; Jun Kikuchi; Hiroshi Ohno

    2015-01-01

    Probiotics are well known as health-promoting agents that modulate intestinal microbiota. However, the molecular mechanisms underlying this effect remain unclear. Using gnotobiotic mice harboring 15 strains of predominant human gut-derived microbiota (HGM), we investigated the effects of Bifidobacterium longum BB536 (BB536-HGM) supplementation on the gut luminal metabolism. Nuclear magnetic resonance (NMR)-based metabolomics showed significantly increased fecal levels of pimelate, a precursor...

  4. Effect of Bifidobacterium longum BB536 yogurt administration on the intestinal environment of healthy adults.

    OpenAIRE

    Ogata, T; Kingaku, M; Yaeshima, T.; Teraguchi, S; Fukuwatari, Y; Ishibashi, N.(Department of Physics, Osaka University, Toyonaka, Osaka 560-0043, Japan); Hayasawa, H.; Fujisawa, T.; Iino, H.

    2011-01-01

    A yogurt supplemented with B. longum BB536 was administered at 250 ml per day for 2 weeks to six healthy volunteers. The effects on the fecal microflora, fecal putrefactive substances, fecal enzymatic activities and fecal properties were examined and compared with the effects of standard yogurt. A significant increase (p<0.05) in the proportion of Bifidobacterium in the fecal microflora was observed following ingestion of yogurt containing B. longum BB536. The numbers of Lactobacillus also...

  5. Influence of whole peptidoglycan of bifidobacterium on cytotoxic effectors produced by mouse peritoneal macrophages

    Institute of Scientific and Technical Information of China (English)

    Li Sheng Wang; Hui Ming Zhu; Dian Yuan Zhou; Yu Lin Wang; Wan Dai Zhang

    2001-01-01

    @@ INTRODUCTION Bifidobacteria are physiologically beneficial bacteria which are perdominant in human intestine ,and possess the most important functions .They play an important role in maintaining microbial balance of the intestine .Furthermore , their presence is thought to be an important indication of health of the body [1-4].Whole peptidoglycan ( WPG) is the major component in the cell wall of bifidobacterium ,which is also a biological responsemodifier with nontoxic side dffcets.

  6. The Bifidobacterium dentium Bd1 genome sequence reflects its genetic adaptation to the human oral cavity.

    Directory of Open Access Journals (Sweden)

    Marco Ventura

    2009-12-01

    Full Text Available Bifidobacteria, one of the relatively dominant components of the human intestinal microbiota, are considered one of the key groups of beneficial intestinal bacteria (probiotic bacteria. However, in addition to health-promoting taxa, the genus Bifidobacterium also includes Bifidobacterium dentium, an opportunistic cariogenic pathogen. The genetic basis for the ability of B. dentium to survive in the oral cavity and contribute to caries development is not understood. The genome of B. dentium Bd1, a strain isolated from dental caries, was sequenced to completion to uncover a single circular 2,636,368 base pair chromosome with 2,143 predicted open reading frames. Annotation of the genome sequence revealed multiple ways in which B. dentium has adapted to the oral environment through specialized nutrient acquisition, defences against antimicrobials, and gene products that increase fitness and competitiveness within the oral niche. B. dentium Bd1 was shown to metabolize a wide variety of carbohydrates, consistent with genome-based predictions, while colonization and persistence factors implicated in tissue adhesion, acid tolerance, and the metabolism of human saliva-derived compounds were also identified. Global transcriptome analysis demonstrated that many of the genes encoding these predicted traits are highly expressed under relevant physiological conditions. This is the first report to identify, through various genomic approaches, specific genetic adaptations of a Bifidobacterium taxon, Bifidobacterium dentium Bd1, to a lifestyle as a cariogenic microorganism in the oral cavity. In silico analysis and comparative genomic hybridization experiments clearly reveal a high level of genome conservation among various B. dentium strains. The data indicate that the genome of this opportunistic cariogen has evolved through a very limited number of horizontal gene acquisition events, highlighting the narrow boundaries that separate commensals from

  7. Protection against Influenza Virus Infection of Mice Fed Bifidobacterium breve YIT4064

    OpenAIRE

    1999-01-01

    Mice fed Bifidobacterium breve YIT4064 and immunized orally with influenza virus were more strongly protected against influenza virus infection of the lower respiratory tract than ones immunized with influenza virus only. The number of mice with enhanced anti-influenza virus immunoglobulin G (IgG) in serum upon oral administration of B. breve YIT4064 and oral immunization with influenza virus was significantly greater than that upon oral immunization with influenza...

  8. Metabolismo di oligosaccaridi prebiotici in Bifidobacterium per il potenziale sviluppo di nuovi prodotti alimentari funzionali

    OpenAIRE

    Zanoni, Simona

    2008-01-01

    The growth and the metabolism of Bifidobacterium adolescentis MB 239 fermenting GOS, lactose, galactose, and glucose were investigated. An unstructerd unsegregated model for growth of B. adolescentis MB 239 in batch cultures was developed and kinetic parameters were calculated with a Matlab algorithm. Galactose was the best carbon source; lactose and GOS led to lower growth rate and cellular yield, but glucose was the poorest carbon source. Lactate, acetate and ethanol yields a...

  9. Alterations in Fecal Lactobacillus and Bifidobacterium Species in Type 2 Diabetic Patients in Southern China Population

    Directory of Open Access Journals (Sweden)

    Kim-Anne eLê

    2013-01-01

    Full Text Available Background: The connection between gut microbiota and metabolism and its role in the pathogenesis of diabetes are increasingly recognized. The objective of this study was to quantitatively measure Bifidobacterium and Lactobacillus species, members of commensal bacteria found in human gut, in type 2 diabetic patients (T2D patients from Southern China. Methods: Fifty patients with T2D and thirty control individuals of similar BMI were recruited from Southern China. T2D and control subjects were confirmed with both oral glucose tolerance test (OGTT and HbA1c measurements. Bifidobacterium and Lactobacillus species in feces were measured by real-time quantitative PCR. Data were analyzed with STATA 11.0 statistical software.Results: In comparison to control subjects T2D patients had significantly more total Lactobacillus (+18%, L. bugaricus (+13%, L. rhamnosum (+37% and L. acidophilus (+48% (P <0.05. In contrast, T2D patients had less amounts of total Bifidobacteria (-7% and B. adolescentis (-12% (P <0.05. Cluster analysis showed that gut microbiota pattern of T2D patients is characterized by greater numbers of L. rhamnosus and L. acidophillus, together with lesser numbers of B. adolescentis (P <0.05. Conclusion: The gut microflora in T2D patients is characterized by greater numbers of Lactobacillus and lesser numbers of Bifidobacterium species.

  10. Membrane filter method to study the effects of Lactobacillus acidophilus and Bifidobacterium longum on fecal microbiota.

    Science.gov (United States)

    Shimizu, Hidenori; Benno, Yoshimi

    2015-11-01

    A large number of commensal bacteria inhabit the intestinal tract, and interbacterial communication among gut microbiota is thought to occur. In order to analyze symbiotic relationships between probiotic strains and the gut microbiota, a ring with a membrane filter fitted to the bottom was used for in vitro investigations. Test strains comprising probiotic nitto strains (Lactobacillus acidophilus NT and Bifidobacterium longum NT) and type strains (L. acidophilus JCM1132(T) and B. longum JCM1217(T) ) were obtained from diluted fecal samples using the membrane filter to simulate interbacterial communication. Bifidobacterium spp., Streptococcus pasteurianus, Collinsella aerofaciens, and Clostridium spp. were the most abundant gut bacteria detected before coculture with the test strains. Results of the coculture experiments indicated that the test strains significantly promote the growth of Ruminococcus gnavus, Ruminococcus torques, and Veillonella spp. and inhibit the growth of Sutterella wadsworthensis. Differences in the relative abundances of gut bacterial strains were furthermore observed after coculture of the fecal samples with each test strain. Bifidobacterium spp., which was detected as the dominant strain in the fecal samples, was found to be unaffected by coculture with the test strains. In the present study, interbacterial communication using bacterial metabolites between the test strains and the gut microbiota was demonstrated by the coculture technique. The detailed mechanisms and effects of the complex interbacterial communications that occur among the gut microbiota are, however, still unclear. Further investigation of these relationships by coculture of several fecal samples with probiotic strains is urgently required.

  11. Bacteriocinogenic Lactococcus lactis subsp. lactis DF04Mi isolated from goat milk: Application in the control of Listeria monocytogenes in fresh Minas-type goat cheese

    Directory of Open Access Journals (Sweden)

    Danielle N. Furtado

    2015-03-01

    Full Text Available Listeria monocytogenes is a pathogen frequently found in dairy products. Its control in fresh cheeses is difficult, due to the psychrotrophic properties and salt tolerance. Bacteriocinogenic lactic acid bacteria (LAB with proven in vitro antilisterial activity can be an innovative technological approach but their application needs to be evaluated by means of in situ tests. In this study, a novel bacteriocinogenic Lactococcus lactis strain (Lc. lactis DF4Mi, isolated from raw goat milk, was tested for control of growth of L. monocytogenes in artificially contaminated fresh Minas type goat cheese during storage under refrigeration. A bacteriostatic effect was achieved, and counts after 10 days were 3 log lower than in control cheeses with no added LAB. However, this effect did not differ significantly from that obtained with a non-bacteriocinogenic Lc. lactis strain. Addition of nisin (12.5 mg/kg caused a rapid decrease in the number of viable L. monocytogenes in the cheeses, suggesting that further studies with the purified bacteriocin DF4Mi may open new possibilities for this strain as biopreservative in dairy products.

  12. Bacteriocinogenic Lactococcus lactis subsp. lactis DF04Mi isolated from goat milk: Application in the control of Listeria monocytogenes in fresh Minas-type goat cheese.

    Science.gov (United States)

    Furtado, Danielle N; Todorov, Svetoslav D; Landgraf, Mariza; Destro, Maria T; Franco, Bernadette D G M

    2015-03-01

    Listeria monocytogenes is a pathogen frequently found in dairy products. Its control in fresh cheeses is difficult, due to the psychrotrophic properties and salt tolerance. Bacteriocinogenic lactic acid bacteria (LAB) with proven in vitro antilisterial activity can be an innovative technological approach but their application needs to be evaluated by means of in situ tests. In this study, a novel bacteriocinogenic Lactococcus lactis strain ( Lc . lactis DF4Mi), isolated from raw goat milk, was tested for control of growth of L. monocytogenes in artificially contaminated fresh Minas type goat cheese during storage under refrigeration. A bacteriostatic effect was achieved, and counts after 10 days were 3 log lower than in control cheeses with no added LAB. However, this effect did not differ significantly from that obtained with a non-bacteriocinogenic Lc. lactis strain. Addition of nisin (12.5 mg/kg) caused a rapid decrease in the number of viable L. monocytogenes in the cheeses, suggesting that further studies with the purified bacteriocin DF4Mi may open new possibilities for this strain as biopreservative in dairy products.

  13. Antitumor effect of cytosine deaminase/5-fluorocytosine suicide gene therapy system mediated by Bifidobacterium infantis on melanoma

    Institute of Scientific and Technical Information of China (English)

    Cheng YI; Ying HUANG; Zhi-ying GUO; Shu-ren WANG

    2005-01-01

    Aim: To construct a Bifidobacterium infantis/CD targeting gene therapy system and observe the antitumor effect of cytosine deaminase/5-fluorocytosine (CD/5-FC) suicide gene therapy system mediated by Bifidobacterium infantis on melanoma in vitro and in vivo. Methods: A recombinant CD/pGEX- 1LamdaT plasmid was transfected into Bifidobacterium infantis by electroporation. Bifidobac terium infantis transfected by recombinant CD/pGEX-1LamdaT plasmid was in cubated with 5-FC anaerobically. Then the supernatant fluid was collected and added to melanoma B16-F10 cells to observe the killing effect for B16-F10 cells.Mice were inoculated with melanoma B 16-F10 cells to establish animal models.The mice were then injected with 5-FC and Bifidobacterium infantis transfected by recombinant CD/pGEX-1LamdaT plasmid. Results:Two segments of approxi mate 4.9 kb and 1.3 kb were extracted from the 6.2 kb recombinant plasmid, which were equal to the size of the pGEX-1LamdaT plasmid and CD gene, respectively.Sequencing results showed that the full length and sequence of nucleotide acid of the inserted gene in extracted recombinant plasmid was completely identical to the CD gene. In vitro, B 16-F10 cells treated by supernatant fluid were remarkably damaged morphologically, and the cell growth was significantly inhibited. Experi ments on the mice melanoma model showed that after treatment with a combination of transfected Bifidobacterium infantis and 5-FC, the tumor volume was significantly inhibited compared with controls. Conclusion: The foreign gene,CD gene, was correctly inserted into pGEX-1LambdaT plasmid and transferred into Bifidobacterium infantis. CD/5-FC suicide gene therapy system mediated by Bifidobacterium infantis demonstrated a good antitumor effect on melanoma in vitro and in vivo.

  14. Physiological responses of Lactococcus lactis ML3 to alternating conditions of growth and starvation

    NARCIS (Netherlands)

    Kunji, E.R.S.; Ubbink, T.; Matin, A.; Poolman, B.; Konings, W.N.

    1993-01-01

    Lactococcus lactis species can survive periods of carbohydrate starvation for relatively long periods of time. In the first hours of starvation, however, the maximal glycolytic and arginine deiminase (ADI) pathway activities decline rapidly. The rate of decrease of the pathway activities diminishes

  15. Genome-Wide Transcriptional Responses to Carbon Starvation in Nongrowing Lactococcus lactis

    NARCIS (Netherlands)

    Ercan,O.; Wels, M.; Smid. E.J.; Kleerebezem, M.

    2015-01-01

    This paper describes the transcriptional adaptations of nongrowing, retentostat cultures of Lactococcus lactis to starvation. Near-zero-growth cultures (μ = 0.0001 h−1) obtained by extended retentostat cultivation were exposed to starvation by termination of the medium supply for 24 h, followed by a

  16. Lactococcus lactis Uses MscL as Its Principal Mechanosensitive Channel

    NARCIS (Netherlands)

    Folgering, Joost H.A.; Moe, Paul C.; Schuurman-Wolters, Gea K.; Blount, Paul; Poolman, Bert

    2005-01-01

    The functions of the mechanosensitive channels from Lactococcus lactis were determined by biochemical, physiological, and electrophysiological methods. Patchclamp studies showed that the genes yncB and mscL encode MscS and MscL-like channels, respectively, when expressed in Escherichia coli or if th

  17. A Case of Infective Endocarditis and Pulmonary Septic Emboli Caused by Lactococcus lactis

    Science.gov (United States)

    Habib, Adib; Asli, Nazih; Geffen, Yuval; Miron, Dan; Elias, Nael

    2016-01-01

    Infective endocarditis is a rare condition in children with normal hearts. We present here a case of previously healthy eleven-year-old girl with infective endocarditis and pulmonary septic emboli caused by a very rare bacterial etiology (Lactococcus lactis). Identification of this pathogen was only made by polymerase chain reaction.

  18. The cmbT gene encodes a novel major facilitator multidrug resistance transporter in Lactococcus lactis.

    Science.gov (United States)

    Filipic, Brankica; Golic, Natasa; Jovcic, Branko; Tolinacki, Maja; Bay, Denice C; Turner, Raymond J; Antic-Stankovic, Jelena; Kojic, Milan; Topisirovic, Ljubisa

    2013-01-01

    Functional characterization of the multidrug resistance CmbT transporter was performed in Lactococcus lactis. The cmbT gene is predicted to encode an efflux protein homologous to the multidrug resistance major facilitator superfamily. The cmbT gene (1377 bp) was cloned and overexpressed in L. lactis NZ9000. Results from cell growth studies revealed that the CmbT protein has an effect on host cell resistance to lincomycin, cholate, sulbactam, ethidium bromide, Hoechst 33342, sulfadiazine, streptomycin, rifampicin, puromycin and sulfametoxazole. Moreover, in vivo transport assays showed that overexpressed CmbT-mediated extrusion of ethidium bromide and Hoechst 33342 was higher than in the control L. lactis NZ9000 strain. CmbT-mediated extrusion of Hoechst 33342 was inhibited by the ionophores nigericin and valinomycin known to dissipate proton motive force. This indicates that CmbT-mediated extrusion is based on a drug-proton antiport mechanism. Taking together results obtained in this study, it can be concluded that CmbT is a novel major facilitator multidrug resistance transporter candidate in L. lactis, with a possible signaling role in sulfur metabolism.

  19. The major facilitator superfamily transporter Knq1p modulates boron homeostasis in Kluyveromyces lactis.

    Science.gov (United States)

    Svrbicka, Alexandra; Toth Hervay, Nora; Gbelska, Yvetta

    2016-03-01

    Boron is an essential micronutrient for living cells, yet its excess causes toxicity. To date, the mechanisms of boron toxicity are poorly understood. Recently, the ScATR1 gene has been identified encoding the main boron efflux pump in Saccharomyces cerevisiae. In this study, we analyzed the ScATR1 ortholog in Kluyveromyces lactis--the KNQ1 gene, to understand whether it participates in boron stress tolerance. We found that the KNQ1 gene, encoding a permease belonging to the major facilitator superfamily, is required for K. lactis boron tolerance. Deletion of the KNQ1 gene led to boron sensitivity and its overexpression increased K. lactis boron tolerance. The KNQ1 expression was induced by boron and the intracellular boron concentration was controlled by Knq1p. The KNQ1 promoter contains two putative binding motifs for the AP-1-like transcription factor KlYap1p playing a central role in oxidative stress defense. Our results indicate that the induction of the KNQ1 expression requires the presence of KlYap1p and that Knq1p like its ortholog ScAtr1p in S. cerevisiae functions as a boron efflux pump providing boron resistance in K. lactis.

  20. Development, molecular characterization and exploitation of the nisin controlled expression system in Lactococcus lactis.

    NARCIS (Netherlands)

    Ruyter, de P.G.G.A.

    1998-01-01

    Lactic acid bacteria are gram-positive bacteria that are widely used in a variety of dairy fermentation processes. Notably, strains of the lactic acid starter bacterium Lactococcus lactis are of great economic importance because of their world-wide use in cheese making. The characteristic aroma, fla

  1. Properties of Nisin Z and Distribution of Its Gene, nisZ, in Lactococcus lactis

    NARCIS (Netherlands)

    Vos, Willem M. de; Mulders, John W.M.; Siezen, Roland J.; Hugenholtz, Jeroen; Kuipers, Oscar P.

    1993-01-01

    Two natural variants of the lantibiotic nisin that are produced by Lactococcus lactis are known. They have a similar structure but differ in a single amino acid residue at position 27: histidine in nisin A and asparagine in nisin Z. The nisin variants were purified to apparent homogeneity, and their

  2. A System To Generate Chromosomal Mutations in Lactococcus lactis Which Allows Fast Analysis of Targeted Genes

    NARCIS (Netherlands)

    Law, Jean; Buist, Girbe; Haandrikman, Alfred; Kok, Jan; Venema, Gerhardus; Leenhouts, Kees

    1995-01-01

    A system for generating chromosomal insertions in lactococci is described. It is based on the conditional replication of lactococcal pWV01-derived Ori+ RepA- vector pORI19, containing lacZα and the multiple cloning site of pUC19. Chromosomal AluI fragments of Lactococcus lactis were cloned in pORI19

  3. Functional Analysis of Promoters in the Nisin Gene Cluster of Lactococcus lactis

    NARCIS (Netherlands)

    Ruyter, Pascalle G.G.A. de; Kuipers, Oscar P.; Beerthuyzen, Marke M.; Alen-Boerrigter, Ingrid van; Vos, Willem M. de

    1996-01-01

    The promoters in the nisin gene cluster nisABTCIPRKFEG of Lactococcus lactis were characterized by primer extension and transcriptional fusions to the Escherichia coli promoterless β-glucuronidase gene (gusA). Three promoters preceding the nisA, nisR, and nisF genes, which all give rise to gusA expr

  4. Rerouting Citrate Metabolism in Lactococcus lactis to Citrate-Driven Transamination

    NARCIS (Netherlands)

    Pudlik, Agata M.; Lolkema, Juke S.

    2012-01-01

    Oxaloacetate is an intermediate of the citrate fermentation pathway that accumulates in the cytoplasm of Lactococcus lactis ILCitM(pFL3) at a high concentration due to the inactivation of oxaloacetate decarboxylase. An excess of toxic oxaloacetate is excreted into the medium in exchange for citrate

  5. Lactococcus lactis Dihydroorotate Dehydrogenase A Mutants Reveal Important Facets of the Enzymatic Function

    DEFF Research Database (Denmark)

    Nørager, Sofie Charlotte; Arent, S; Björnberg, Olof

    2003-01-01

    and 1B, and class 2. This division corresponds to differences in cellular location and the nature of the electron acceptor. Herein we report a study of Lactococcus lactis DHODA, a representative of the class 1A enzymes. Based on the DHODA structure we selected seven residues that are highly conserved...

  6. Transcriptome analysis of the Lactococcus lactis ArgR and AhrC regulons

    DEFF Research Database (Denmark)

    Larsen, Rasmus; van Hijum, Sacha A. F. T.; Martinussen, Jan;

    2008-01-01

    In previous studies, we have shown that direct protein-protein. interaction between the two regulators ArgR and AhrC in Lactococcus lactis is required for arginine-dependent repression of the biosynthetic argC promoter and the activation of the catabolic arcA promoter. Here, we establish the global...

  7. Adaptation of Lactococcus lactis to high growth temperature leads to a dramatic increase in acidification rate

    DEFF Research Database (Denmark)

    Chen, Jun; Shen, Jing; Hellgren, Lars;

    2015-01-01

    Lactococcus lactis is essential for most cheese making, and this mesophilic bacterium has its growth optimum around 30 °C. We have, through adaptive evolution, isolated a mutant TM29 that grows well up to 39 °C, and continuous growth at 40 °C is possible if pre-incubated at a slightly lower...

  8. Characterization of the Lactococcus lactis lactose genes and regulation of their expression.

    NARCIS (Netherlands)

    Rooijen, van R.J.

    1993-01-01

    An important trait of the lactic acid bacterium Lactococcus lactis , that is used in industrial dairy fermentations, is the conversion of lactose into lactic acid. The enzymatic steps involved in the breakdown of lactose, that is transported into the cell via a phosphoenolpyruvate-dependent lactose

  9. Immunogenicity of a malaria parasite antigen displayed by Lactococcus lactis in oral immunisations

    NARCIS (Netherlands)

    Ramasamy, R; Yasawardena, S; Zomer, A; Venema, G; Kok, J; Leenhouts, K

    2006-01-01

    A putative protective protein from Plasmodium falciparum merozoites, MSA2, was expressed in two different ways on the cell surface of the Gram-positive food-grade bacterium, Lactococcus lactis. The first display format exploits an LPXTG-type anchoring motif of the lactococcal proteinase PrtP to cova

  10. Comparative analyses of prophage-like elements present in two Lactococcus lactis strains

    NARCIS (Netherlands)

    Ventura, Marco; Zomer, Aldert; Canchaya, Carlos; O'Connell-Motherway, Mary; Kuipers, Oscar; Turroni, Francesca; Ribbera, Angela; Foroni, Elena; Buist, Girbe; Wegmann, Udo; Shearman, Claire; Gasson, Michael J.; Fitzgerald, Gerald F.; Kok, Jan; van Sinderen, Douwe; O’Connell-Motherway, Mary

    2007-01-01

    In this study, we describe the genetic organizations of six and five apparent prophage-like elements present in the genomes of the Lactococcus lactis subsp. cremoris strains MG1363 and SK11, respectively. Phylogenetic investigation as well bioinformatic analyses indicates that all 11 prophages belon

  11. Kinetics and regulation of lactose transport and metabolism in Kluyveromyces lactis JA6.

    Science.gov (United States)

    Santos, A M; Silveira, W B; Fietto, L G; Brandão, R L; Castro, I M

    2014-07-01

    Kluyveromyces lactis strains are able to assimilate lactose. They have been used industrially to eliminate this sugar from cheese whey and in other industrial products. In this study, we investigated specific features and the kinetic parameters of the lactose transport system in K. lactis JA6. In lactose grown cells, lactose was transported by a system transport with a half-saturation constant (K s) of 1.49 ± 0.38 mM and a maximum velocity (V max) of 0.96 ± 0.12 mmol. (g dry weight)(-1) h(-1) for lactose. The transport system was constitutive and energy-dependent. Results obtained by different approaches showed that the lactose transport system was regulated by glucose at the transcriptional level and by glucose and other sugars at a post-translational level. In K. lactis JA6, galactose metabolization was under glucose control. These findings indicated that the regulation of lactose-galactose regulon in K. lactis was similar to the regulation of galactose regulon in Saccharomyces cerevisiae.

  12. Carbon catabolite repression and global control of the carbohydrate metabolism in Lactococcus lactis.

    NARCIS (Netherlands)

    Luesink, E.J.

    1998-01-01

    In view of the economic importance of fermented dairy products considerable scientific attention has been given to various steps of fermentation processes, including the L-lactate formation of lactic acid bacteria (de Vos, 1996). In particular, the carbohydrate metabolism of L. lactis has been the s

  13. Dependence of Streptococcus lactis Phosphate Transport on Internal Phosphate Concentration and Internal pH

    NARCIS (Netherlands)

    POOLMAN, B; NIJSSEN, RMJ; KONINGS, WN

    1987-01-01

    Uptake of phosphate by Streptococcus lactis ML3 proceeds in the absence of a proton motive force, but requires the synthesis of ATP by either arginine or lactose metabolism. The appearance of free Pi internally in arginine-metabolizing cells corresponded quantitatively with the disappearance of extr

  14. Topology of LcnD, a protein implicated in the transport of bacteriocins from Lactococcus lactis

    NARCIS (Netherlands)

    Franke, Christian M.; Leenhouts, Kees J.; Haandrikman, Alfred J.; Kok, Jan; Venema, Gerard; Venema, Koen

    1996-01-01

    Four in-frame translational fusions to both the reporter proteins beta-galactosidase and alkaline phosphatase support a topological model of LcnD, a protein implicated in the transport of several bacteriocins from Lactococcus lactis, in which the N-terminal part is located intracellularly and one tr

  15. Quantitative physiology of Lactococcus lactis at extreme low-growth rates

    NARCIS (Netherlands)

    Ercan, O.; Smid, E.J.; Kleerebezem, M.

    2013-01-01

    This paper describes the metabolic adaptation of Lactococcus lactis during the transition from a growing to a non-growing state using retentostat cultivation. Under retentostat cultivation, the specific growth rate decreased from 0.025 h-1 to 0.0001 h-1 in 42 days, while doubling time increased to m

  16. Genome-wide transcriptional responses to carbon starvation in nongrowing Lactococcus lactis

    NARCIS (Netherlands)

    Ercan, O.; Wels, M.; Smid, E.J.; Kleerebezem, M.

    2015-01-01

    This paper describes the transcriptional adaptations of nongrowing, retentostat cultures of Lactococcus lactis to starvation. Near-zero-growth cultures (µ = 0.0001 h-1) obtained by extended retentostat cultivation were exposed to starvation by termination of the medium supply for 24 h, followed by a

  17. Physiological and molecular adaptations of Lactococcus lactis to near-zero growth conditions

    NARCIS (Netherlands)

    Ercan, O.

    2014-01-01

    Lactococcus lactis is an important lactic acid bacteria (LAB) species that is used for the manufacture of dairy products, such as cheese, buttermilk, and other fermented products. The predominant function of this bacterium in dairy fermentation is the production of lactic acid, as its major fermenta

  18. A possible contribution of mRNA secondary structure to translation initiation efficiency in Lactococcus lactis

    NARCIS (Netherlands)

    Guchte, Maarten van de; Lende, Ted van der; Kok, Jan; Venema, Gerard

    1991-01-01

    Gene expression signals derived from Lactococcus lactis were linked to lacZ-fused genes with different 5'-nucleotide sequences. Computer predictions of mRNA secondary structure were combined with lacZ expression studies to direct base-substitutions that could possibly influence gene expression. Muta

  19. Targeting diseases with genetically engineered Lactococcus lactis and its course towards medical translation

    NARCIS (Netherlands)

    Villatoro-Hernandez, Julio; Montes-de-Oca-Luna, Roberto; Kuipers, Oscar P.

    2011-01-01

    The use of the lactic acid bacterium Lactococcus lactis, primarily used in food fermentations, as therapeutic agent is no longer speculative but an imminent reality. After the successful completion of Phase I and II clinical trials in humans for the treatment of inflammatory bowel disease, an ongoin

  20. Insertion-Sequence-Mediated Mutations Isolated During Adaptation to Growth and Starvation in Lactococcus lactis.

    NARCIS (Netherlands)

    Visser, de J.A.G.M.; Akkermans, A.D.L.; Hoekstra, R.F.; Vos, de W.M.

    2004-01-01

    We studied the activity of three multicopy insertion sequence (IS) elements in 12 populations of Lactococcus lactis IL1403 that evolved in the laboratory for 1000 generations under various environmental conditions (growth or starvation and shaken or stationary). Using RFLP analysis of single-clone r

  1. Systematic identification of tRNAome and its dynamics in Lactococcus lactis

    NARCIS (Netherlands)

    Puri, Pranav; Wetzel, Collin; Saffert, Paul; Gaston, Kirk W.; Russel, Susan P.; Cordero Varela, Juan A.; van der Vlies, Pieter; Zhang, Gong; Limbach, Patrick A.; Ignatova, Zoya; Poolman, Bert

    2014-01-01

    Transfer RNAs (tRNAs) through their abundance and modification pattern significantly influence protein translation. Here, we present a systematic analysis of the tRNAome of Lactococcus lactis. Using the next-generation sequencing approach, we identified 40 tRNAs which carry 16 different post-transcr

  2. Microbial domestication signatures of Lactococcus lactis can be reproduced by experimental evolution

    NARCIS (Netherlands)

    Bachmann, H.; Starrenburg, M.J.C.; Molenaar, D.; Kleerebezem, M.; Hylckama Vlieg, van J.E.T.

    2012-01-01

    Experimental evolution is a powerful approach to unravel how selective forces shape microbial genotypes and phenotypes. To this date, the available examples focus on the adaptation to conditions specific to the laboratory. The lactic acid bacterium Lactococcus lactis naturally occurs on plants and i

  3. CONTINUOUS MEASUREMENT OF THE CYTOPLASMIC PH IN LACTOCOCCUS-LACTIS WITH A FLUORESCENT PH INDICATOR

    NARCIS (Netherlands)

    MOLENAAR, D; ABEE, T; KONINGS, WN

    1991-01-01

    The cytoplasmic pH of Lactococcus lactis was studied with the fluorescent pH indicator 2',7'-bis-(2-carboxyethyl)-5 (and-6)-carboxyfluorescein (BCECF). A novel method was applied for loading bacterial cells with BCECF, which consists of briefly treating a dense cell suspension with acid in the prese

  4. A comparative study between inhibitory effect of L. lactis and nisin on important pathogenic bacteria in Iranian UF Feta cheese

    Directory of Open Access Journals (Sweden)

    Saeed Mirdamadi

    2015-02-01

    Full Text Available   Introduction : In the present study, the inhibitory effect of nisin-producing Lactococcus lactis during co-culture and pure standard nisin were assessed against selected foodborne pathogenes in growth medium and Iranian UF Feta cheese. In comparison L lactis, not only proves flavor but also plays a better role in microbial quality of Iranian UF Feta cheese as a model of fermented dairy products.   Materials and method s: L. lactis subsp. lactis as nisin producer strain, Listeria monocytogenes, Escherichia coli and Staphylococcus aureus as pathogenic strains were inoculated in Ultra-Filtered Feta cheese. Growth curve of bacterial strains were studied by colony count method in growth medium and UF Feta cheese separately and during co-culture with L. lactis. Nisin production was determined by agar diffusion assay method against susceptible test strain and confirmed by RP-HPLC analysis method.   Results : Counts of L. monocytogenes decreased in cheese sample containing L. lactis and standard nisin, to 103 CFU/g after 7 days and it reached to undetectable level within 2 weeks. S. aureus counts remained at its initial number, 105 CFU/g, after 7 days then decreased to 104 CFU/g on day 14 and it was not detectable on day 28. E. coli numbers increased in both treatments after 7 days and then decreased to 104 CFU/g after 28 days. Despite the increasing number of E. coli in growth medium containing nisin, due to the synergistic effect of nisin and other metabolites produced by Lactococcus lactis and starter cultures, the number of E. coli decreased with slow rate . Discussion and conclusion : The results showed, L. monocytogenes was inhibited by L. lactis before entering the logarithmic phase during co-culture. S. aureus was also inhibited during co-culture, but it showed less sensitivity in comparison with L. monocytogenes. However, the number of E. coli remained steady in co-culture with L. lactis. Also, we found that, in all cheese samples, E

  5. Effect of Pre-Stressing on the Acid-Stress Response in Bifidobacterium Revealed Using Proteomic and Physiological Approaches.

    Directory of Open Access Journals (Sweden)

    Junhua Jin

    Full Text Available Weak acid resistance limits the application of Bifidobacteria as a probiotic in food. The acid tolerance response (ATR, caused by pre-stressing cells at a sublethal pH, could improve the acid resistance of Bifidobacteria to subsequent acid stress. In this study, we used Bifidobacterium longum sub. longum BBMN68 to investigate the effect of the ATR on the acid stress response (ASR, and compared the difference between the ATR and the ASR by analyzing the two-dimensional-PAGE protein profiles and performing physiological tests. The results revealed that a greater abundance of proteins involved in carbohydrate metabolism and protein protection was present after the ASR than after the ATR in Bifidobacterium. Pre-stressing cells increased the abundance of proteins involved in energy production, amino acid metabolism, and peptidoglycan synthesis during the ASR of Bifidobacterium. Moreover, after the ASR, the content of ATP, NH3, thiols, and peptidoglycan, the activity of H+-ATPase, and the maintenance of the intracellular pH in the pre-stressed Bifidobacterium cells was significantly higher than in the uninduced cells. These results provide the first explanation as to why the resistance of Bifidobacterium to acid stress improved after pre-stressing.

  6. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of health claims related to a combination of Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum M 20/5 and Bifidobacterium longum SP 07/3 and maintenance of upper respiratory tract defence

    DEFF Research Database (Denmark)

    Tetens, Inge

    of Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum M 20/5 and Bifidobacterium longum SP 07/3 and maintenance of upper respiratory tract defence against pathogens. The food constituent that is the subject of the claim, a combination of Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum M 20....../5 and Bifidobacterium longum SP 07/3, is sufficiently characterised. The claimed effect, maintenance of upper respiratory tract defence against pathogens, is a beneficial physiological effect. The proposed target population is the general adult population. The Panel notes that the only human intervention study provided...... did not show an effect of consumption of a combination of Lactobacillus gasseri PA 16/8, Bifidobacterium bifidum M 20/5 and Bifidobacterium longum SP 07/3 on defence against pathogens in the upper respiratory tract. On the basis of the data presented, the Panel concludes that a cause and effect...

  7. Expression of mptC of Listeria monocytogenes induces sensitivity to class IIa bacteriocins in Lactococcus lactis.

    Science.gov (United States)

    Ramnath, Manilduth; Arous, Safia; Gravesen, Anne; Hastings, John W; Héchard, Yann

    2004-08-01

    Sensitivity to class IIa bacteriocins from lactic acid bacteria was recently associated with the mannose phosphotransferase system (PTS) permease, in Listeria monocytogenes. To assess the involvement of this protein complex in class IIa bacteriocin activity, the mptACD operon, encoding, was heterologously expressed in an insensitive species, namely Lactococcus lactis, using the NICE double plasmid system. Upon induction of the cloned operon, the recombinant Lc. lactis became sensitive to leucocin A. Pediocin PA-1 and enterocin A also showed inhibitory activity against Lc. lactis cultures expressing mptACD. Furthermore, the role of the three genes of the mptACD operon was investigated. Derivative plasmids containing various combinations of these three genes were made from the parental mptACD plasmid by divergent PCR. The results showed that expression of mptC alone is sufficient to confer sensitivity to class IIa bacteriocins in Lc. lactis.

  8. Powerful methods to establish chromosomal markers in Lactococcus lactis: an analysis of pyrimidine salvage pathway mutants obtained by positive selections

    DEFF Research Database (Denmark)

    Martinussen, Jan; Hammer, Karin

    1995-01-01

    phosphoribosyltransferase (upp), uridindcytidine kinase (udk), pyrimidine nucleoside phosphorylase (pdp), cytidine/deoxycytidine deaminase (dd), thymidine kinase (tdk) and purine nucleoride phosphorylase (pup). Based on an analysis of the mutants obtained, the pathways by which L. lactis metabolizes uracil...

  9. Analysis of infant isolates of Bifidobacterium breve by comparative genome hybridization indicates the existence of new subspecies with marked infant specificity

    NARCIS (Netherlands)

    Boesten, R.; Schuren, F.; Wind, R.D.; Knol, J.; Vos, W.M. de

    2011-01-01

    A total of 20 Bifidobacterium strains were isolated from fecal samples of 4 breast- and bottle-fed infants and all were characterized as Bifidobacterium breve based on 16S rRNA gene sequence and metabolic analysis. These isolates were further characterized and compared to the type strains of B. brev

  10. Production of Recombinant Peanut Allergen Ara h 2 using Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    Frøkiær Hanne

    2007-08-01

    Full Text Available Abstract Background Natural allergen sources can supply large quantities of authentic allergen mixtures for use as immunotherapeutics. However, such extracts are complex, difficult to define, vary from batch to batch, which may lead to unpredictable efficacy and/or unacceptable levels of side effects. The use of recombinant expression systems for allergen production can alleviate some of these issues. Several allergens have been tested in high-level expression systems and in most cases show immunereactivity comparable to their natural counterparts. The gram positive lactic acid bacterium Lactococcus lactis is an attractive microorganism for use in the production of protein therapeutics. L. lactis is considered food grade, free of endotoxins, and is able to secrete the heterologous product together with few other native proteins. Hypersensitivity to peanut represents a serious allergic problem. Some of the major allergens in peanut have been described. However, for therapeutic usage more information about the individual allergenic components is needed. In this paper we report recombinant production of the Ara h 2 peanut allergen using L. lactis. Results A synthetic ara h 2 gene was cloned into an L. lactis expression plasmid containing the P170 promoter and the SP310mut2 signal sequence. Flask cultures grown overnight showed secretion of the 17 kDa Ara h 2 protein. A batch fermentation resulted in 40 mg/L recombinant Ara h 2. Purification of Ara h 2 from the culture supernatant was done by hydrophobic exclusion and size separation. Mass spectrometry and N-terminal analysis showed a recombinant Ara h 2 of full length and correctly processed by the signal peptidase. The immunological activity of recombinant Ara h 2 was analysed by ELISA using antibodies specific for native Ara h 2. The recombinant Ara h 2 showed comparable immunereactivity to that of native Ara h 2. Conclusion Recombinant production of Ara h 2 using L. lactis can offer high yields

  11. Experimental study on producing tomato vinegar by Bifidobacterium%双歧番茄醋的实验研究

    Institute of Scientific and Technical Information of China (English)

    王春耀; 张德纯; 张名均; 李金玲; 刘明方

    2011-01-01

    Objective To use tomato as main material,and study the fermentation process of tomato vinegar by Bifidobacterium and Acetobacter pasteurianus.Method The optimal process was selected by Orthogonal test.Result The final acidlty of Bifidobacterium vinegar was 27 g/L; The total count of Bifidobacterium was 1.9 × 1011 CFU/mL; Viable count of Bifidobacterium was 5.5 × 107 CFU/mL in five days.The product had a brown color and good gloss, with smell of ripe tomato and moderate taste.Conclusion Bifidobacterium and Acetobacter pasteurianus can survive together in tomato juice and it is feasible to use them to produce Bifidobacterium tomato vinegar.%目的 以番茄为主要原料,对双歧杆菌和醋酸杆菌共同发酵研制双歧番茄醋的工艺进行研究.方法 通过正交试验筛选最适工艺.结果 双歧番茄醋的最终醋酸度为27 g/L,含双歧杆菌总菌为1.9×1011 CFU/mL,5 d内双歧杆菌活菌为5.5×107 CFU/mL.双歧番茄醋棕黄色,光泽度好,有成熟番茄香味,入口酸甜适中.结论 双歧杆菌与醋酸杆菌在可以番茄汁中共生,此方法制备双歧番茄醋可行.

  12. Classification of a moderately oxygen-tolerant isolate from baby faeces as Bifidobacterium thermophilum

    Directory of Open Access Journals (Sweden)

    Fliss Ismaïl

    2007-08-01

    Full Text Available Abstract Background Bifidobacteria are found at varying prevalence in human microbiota and seem to play an important role in the human gastrointestinal tract (GIT. Bifidobacteria are highly adapted to the human GIT which is reflected in the genome sequence of a Bifidobacterim longum isolate. The competitiveness against other bacteria is not fully understood yet but may be related to the production of antimicrobial compounds such as bacteriocins. In a previous study, 34 Bifidobacterium isolates have been isolated from baby faeces among which six showed proteinaceous antilisterial activity against Listeria monocytogenes. In this study, one of these isolates, RBL67, was further identified and characterized. Results Bifidobacterium isolate RBL67 was classified and characterized using a polyphasic approach. RBL67 was classified as Bifidobacterium thermophilum based on phenotypic and DNA-DNA hybridization characteristics, although 16S rDNA analyses and partial groEL sequences showed higher homology with B. thermacidophilum subsp. porcinum and B. thermacidophilum subsp. thermacidophilum, respectively. RBL67 was moderately oxygen-tolerant and was able to grow at pH 4 and at a temperature of 47°C. Conclusion In order to assign RBL67 to a species, a polyphasic approach was used. This resulted in the classification of RBL67 as a Bifidobacterium thermophilum strain. To our knowledge, this is the first report about B. thermophilum isolated from baby faeces since the B. thermophilum strains were related to ruminants and swine faeces before. B. thermophilum was previously only isolated from animal sources and was therefore suggested to be used as differential species between animal and human contamination. Our findings may disapprove this suggestion and further studies are now conducted to determine whether B. thermophilum is distributed broader in human faeces. Furthermore, the postulated differentiation between human and animal strains by growth above 45

  13. Bifidobacterium bifidum Actively Changes the Gene Expression Profile Induced by Lactobacillus acidophilus in Murine Dendritic Cells

    DEFF Research Database (Denmark)

    Weiss, Gudrun Margarethe; Rasmussen, Simon; Fink, Lisbeth Nielsen;

    2010-01-01

    Dendritic cells (DC) play a pivotal regulatory role in activation of both the innate as well as the adaptive immune system by responding to environmental microorganisms. We have previously shown that Lactobacillus acidophilus induces a strong production of the pro-inflammatory and Th1 polarizing...... cytokine IL-12 in DC, whereas bifidobacteria do not induce IL-12 but inhibit the IL-12 production induced by lactobacilli. In the present study, genome-wide microarrays were used to investigate the gene expression pattern of murine DC stimulated with Lactobacillus acidophilus NCFM and Bifidobacterium...

  14. Nucleotide Sequence and Analysis of an orotate transporter-containing plasmid isolated from the Lactococcus lactis ssp. lactis biovar diacetylactis strain DB0410

    DEFF Research Database (Denmark)

    Defoor, Els Marie Celine; Martinussen, Jan

    determined. Fifteen open reading frames (ORFs) were encountered of which three insertion-sequence (IS) elements identified as two IS946 and one IS982. Two ORFs are incomplete due to the insertion of an IS element in their carboxy terminal end. Homologs for four ORFs were found on the IL1403 sequence: the cop...... and molecular cloning, we identified the open reading frames on pDBORO necessary for the utilization of orotate as the sole pyrimidine source. Surprisingly, homologs are found on the Lactococcus lactis IL1403 and MG1363 chromosomes despite the fact that they are resistant towards fluoroorotate....

  15. Biosorption of silver cations onto Lactococcus lactis and Lactobacillus casei isolated from dairy products

    Science.gov (United States)

    Milanowski, Maciej; Pomastowski, Paweł; Railean-Plugaru, Viorica; Rafińska, Katarzyna; Ligor, Tomasz; Buszewski, Bogusław

    2017-01-01

    The current work deals with the phenomenon of silver cations uptake by two kinds of bacteria isolated from dairy products. The mechanism of sorption of silver cations by Lactococcus lactis and Lactobacillus casei bacteria was investigated. Inductively coupled plasma–mass spectrometry (ICP-MS) was used for determination of silver concentration sorbed by bacteria. Analysis of charge distribution was conducted by diffraction light scattering method. Changes in the ultrastructure of Lactococcus lactis and Lactobacillus casei cells after treatment with silver cations were investigated using transmission electron microscopy observation. Molecular spectroscopy methods, namely Fourier transform-infrared spectroscopy (FT-IR) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) were employed for description of the sorption mechanism. Moreover, an analysis of volatile organic compounds (VOCs) extracted from bacterial cells was performed. PMID:28362838

  16. Analysis of heat shock gene expression in Lactococcus lactis MG1363

    DEFF Research Database (Denmark)

    Arnau, José; Sørensen, Kim; Appel, Karen Fuglede

    1996-01-01

    The induction of the heat shock response in Lactococcus lactis subsp. cremoris strain MG1363 was analysed at the RNA level using a novel RNA isolation procedure to prevent degradation. Cloning of the dnaJ and groEL homologous was carried out. Nothern blot analysis showed a similar induction pattern......, although maximum induction was observed earlier for orf1 and grpE. Novel transcript sizes were detected in heat-shocked cells. The induction kinetics observed for ftsH suggested a different regulation for this gene. Experimental evidence for a prenounced transcriptional regulation being involved...... in the heat shock response in L. lactis MG1363 is presented. A gene located downstream of the dnaK operon in strain MG1363, named orf4, was shown not to be regulated by heat shock....

  17. Production of recombinant peanut allergen Ara h 2 using Lactococcus lactis

    DEFF Research Database (Denmark)

    Glenting, J.; Poulsen, Lars K.; Kato, K.;

    2007-01-01

    of the major allergens in peanut have been described. However, for therapeutic usage more information about the individual allergenic components is needed. In this paper we report recombinant production of the Ara h 2 peanut allergen using L. lactis. Results: A synthetic ara h 2 gene was cloned into an L....... lactis expression plasmid containing the P170 promoter and the SP310mut2 signal sequence. Flask cultures grown overnight showed secretion of the 17 kDa Ara h 2 protein. A batch fermentation resulted in 40 mg/ L recombinant Ara h 2. Purification of Ara h 2 from the culture supernatant was done...... by hydrophobic exclusion and size separation. Mass spectrometry and N- terminal analysis showed a recombinant Ara h 2 of full length and correctly processed by the signal peptidase. The immunological activity of recombinant Ara h 2 was analysed by ELISA using antibodies specific for native Ara h 2...

  18. Purification and Characterization of a Bacteriocin Produced by Lactobacillus lactis Isolated from Marine Environment

    Directory of Open Access Journals (Sweden)

    P. Manivasagan

    2010-03-01

    Full Text Available Bacteriocin producing Lactobacillus lactis strain isolated from marine environment, showed broadrange of antibacterial activity against some major food borne pathogens. Maximum bacteriocin production wasobserved at 30°C , pH 6.0 and 1.5% sodium chloride solution. In addition of enzymes, "-amylase, DNase,RNase and lipase were slightly positive effect bacteriocin production. Proteinase K and pepsin were stronglyinhibited bacteriocin production. Among detergents, Sodium dodecyl sulphate (SDS, Tween 80 and TritoneX-100 stimulated bacteriocin production and strongly inhibited by EDTA and urea. The bacteriocin has purifiedby ammonium sulphate precipitate and ion exchange (DEAE cellulose chromatography. Biochemically it waspure protein moiety and the molecular weight was 94 kDa. The study revealed the possibility of usingbacteriocin as a food preservative and the L. lactis strain as probiotic.

  19. The pyrimidine operon pyrRPB-carA from Lactococcus lactis

    DEFF Research Database (Denmark)

    Martinussen, Jan; Schallert, J.; Andersen, Birgit;

    2001-01-01

    The four genes pyrR, pyrP, pyrB, and carA were found to constitute an operon in Lactococcus lactis subsp, lactis MG1363. The functions of the different genes were established by mutational analysis. The first gene in the operon is the pyrimidine regulatory gene, pyrR, which is responsible...... for the regulation of the expression of the pyrimidine biosynthetic genes leading to UMP formation. The second gene encodes a membrane-bound high-affinity uracil permease, required for utilization of exogenous uracil. The last two genes in the operon, pyrB and carA, encode pyrimidine biosynthetic enzymes; aspartate....... The expression of the pyrimidine biosynthetic genes including the pyrRPB-carA operon is subject to control at the transcriptional level, most probably by an attenuator mechanism in which PyrR acts as the regulatory protein....

  20. Study of the transgalactosylation activity of ß-galactosidase from a new strain Kluyveromyces lactis 3

    Directory of Open Access Journals (Sweden)

    ILIA ILIEV

    2012-01-01

    Full Text Available Beta-galactosidase (EC.3.2.1.23 is an important enzyme industrially used for the hydrolysis of lactose from milk and milk whey for several applications. Lately, the importance of this enzyme was enhanced by its galactosyltransferase activity, which is responsible for synthesis of transgalactosylated oligosaccharides that act as prebiotics with several beneficial effects on the consumers. ß-Galactosidase production by Kluyveromyces lactis 3 was studied in shake flask culture. The highest enzymatic activity was obtained at 10-th hour of the fermentation. The optimum temperature for transferase activity was 50°C. When incubated with 30% lactose in 50 mM phosphate buffer (pH 6.0 the enzyme can synthesize up to 41% galacto-oligosaccharides (GalOS. β-Galactosidase from strain Kluyveromyces lactis 3 produces mainly oligosaccharides with degree of polymerization (DP 6 at 40°C and with DP 3 at 50°C.

  1. Increased biomass yield of Lactococcus lactis during energetically limited growth and respiratory conditions

    DEFF Research Database (Denmark)

    Købmann, Brian Jensen; Blank, Lars Mathias; Solem, Christian

    2008-01-01

    Lactococcus lactis is known to be capable of respiration under aerobic conditions in the presence of haemin. In the present study the effect of respiration on ATP production during growth on different sugars was examined. With glucose as the sole carbon source, respiratory conditions in L. lactis...... MG1363 resulted in only a minor increase, 21%, in biomass yield. Since ATP production through substrate-level phosphorylation was essentially identical with and without respiration, the increased biomass yield was a result of energy-saving under respiratory conditions estimated to be 0.4 mol of ATP....../mol of glucose. With maltose as the energy source, the increase in biomass yield amounted to 51% compared with an aerobic culture that lacked haemin. This higher ATP yield was obtained by redirecting pyruvate metabolism from lactate to acetate production, and from savings through respiration. However, even after...

  2. The Pasteur effect and catabolite repression in an oxidative yeast, Kluyveromyces lactis.

    Science.gov (United States)

    Royt, P W; MacQuillan, A M

    1979-01-01

    The presence of the Pasteur effect in Kluyveromyces lactis grown in glucose was shown by azide-stimulated glucose fermentation. Extracts from these cells contained ATP-sensitive phosphofructokinase activity. Cells grown on succinate oxidized glucose slowly at first without azide-stimulated rates of fermentation. Phosphofructokinase in these cells was ATP-insensitive. The activity of NAD+-isocitrate dehydrogenase in cell extracts did not require AMP activation. These results suggested the presence of a Pasteur effect in glucose-grown but not in succinate-grown K. lactis, mediated by (a) ATP inhibition of phosphofructokinase (b) possibly via feedback control of glucose transport, but not by AMP activation of isocitrate dehydrogenase. Azide inhibition of the Pasteur effect during growth of the cells did not lead to catabolite repression of respiratory activity. The results therefore suggest that the Pasteur effect does not inhibit the development of a Crabtree effect in oxidative yeasts.

  3. PpiA, a surface PPIase of the cyclophilin family in Lactococcus lactis.

    Directory of Open Access Journals (Sweden)

    Nicolas Trémillon

    Full Text Available BACKGROUND: Protein folding in the envelope is a crucial limiting step of protein export and secretion. In order to better understand this process in Lactococcus lactis, a lactic acid bacterium, genes encoding putative exported folding factors like Peptidyl Prolyl Isomerases (PPIases were searched for in lactococcal genomes. RESULTS: In L. lactis, a new putative membrane PPIase of the cyclophilin subfamily, PpiA, was identified and characterized. ppiA gene was found to be constitutively expressed under normal and stress (heat shock, H(2O(2 conditions. Under normal conditions, PpiA protein was synthesized and released from intact cells by an exogenously added protease, showing that it was exposed at the cell surface. No obvious phenotype could be associated to a ppiA mutant strain under several laboratory conditions including stress conditions, except a very low sensitivity to H(2O(2. Induction of a ppiA copy provided in trans had no effect i on the thermosensitivity of an mutant strain deficient for the lactococcal surface protease HtrA and ii on the secretion and stability on four exported proteins (a highly degraded hybrid protein and three heterologous secreted proteins in an otherwise wild-type strain background. However, a recombinant soluble form of PpiA that had been produced and secreted in L. lactis and purified from a culture supernatant displayed both PPIase and chaperone activities. CONCLUSIONS: Although L. lactis PpiA, a protein produced and exposed at the cell surface under normal conditions, displayed a very moderate role in vivo, it was found, as a recombinant soluble form, to be endowed with folding activities in vitro.

  4. Physiological and regulatory effects of controlled overproduction of five cold shock proteins of Lactococcus lactis

    NARCIS (Netherlands)

    Wouters, J.A.; Mailhes, M.; Rombouts, F.M.; Vos, de W.M.; Kuipers, O.P.; Abee, T.

    2000-01-01

    The physiological and regulatory effects of overproduction of five cold shock proteins (CSPs) of Lactococcus lactis were studied. CspB, CspD, and CspE could be overproduced at high levels (up to 19␘f the total protein), whereas for CspA and CspC limited overproduction (0.3 to 0.5␘f the total protein

  5. In vivo imaging of Lactococcus lactis, Lactobacillus plantarum and Escherichiacoli expressing infrared fluorescent protein in mice

    OpenAIRE

    Berlec, Aleš; Štrukelj, Borut; Završnik, Janja; Turk, Boris; Butinar, Miha

    2016-01-01

    Background In vivo imaging of orally administered lactic acid bacteria (LAB) and commensal bacteria in mice is shown to provide information on the spatial and temporal distribution of bacteria in the gastrointestinal tract. The bacteria can be detected and monitored using bioluminescence or near-infrared fluorescence. Results Fluorescence imaging of bacteria was established by expressing the infrared fluorescent protein IRFP713 in Lactococcus lactis, Lactobacillus plantarum and Escherichia co...

  6. Two Lactococcus lactis thioredoxin paralogues play different roles in responses to arsenate and oxidative stress

    DEFF Research Database (Denmark)

    Efler, Petr; Kilstrup, Mogens; Johnsen, Stig;

    2015-01-01

    Thioredoxin (Trx) maintains intracellular thiol groups in a reduced state and is involved in a wide range of cellular processes, including ribonucleotide reduction, sulphur assimilation, oxidative stress responses and arsenate detoxification. The industrially important lactic acid bacterium Lacto...... the phenotype of the ΔtrxA mutant matches established functions of WCGPC-type Trx while TrxD appears to play a more restricted role in stress resistance of Lac. lactis....

  7. Physiological adaptation of the bacterium Lactococcus lactis in response to the production of human CFTR.

    Science.gov (United States)

    Steen, Anton; Wiederhold, Elena; Gandhi, Tejas; Breitling, Rainer; Slotboom, Dirk Jan

    2011-07-01

    Biochemical and biophysical characterization of CFTR (the cystic fibrosis transmembrane conductance regulator) is thwarted by difficulties to obtain sufficient quantities of correctly folded and functional protein. Here we have produced human CFTR in the prokaryotic expression host Lactococcus lactis. The full-length protein was detected in the membrane of the bacterium, but the yields were too low (proteins) for in vitro functional and structural characterization, and induction of the expression of CFTR resulted in growth arrest. We used isobaric tagging for relative and absolute quantitation based quantitative proteomics to find out why production of CFTR in L. lactis was problematic. Protein abundances in membrane and soluble fractions were monitored as a function of induction time, both in CFTR expression cells and in control cells that did not express CFTR. Eight hundred and forty six proteins were identified and quantified (35% of the predicted proteome), including 163 integral membrane proteins. Expression of CFTR resulted in an increase in abundance of stress-related proteins (e.g. heat-shock and cell envelope stress), indicating the presence of misfolded proteins in the membrane. In contrast to the reported consequences of membrane protein overexpression in Escherichia coli, there were no indications that the membrane protein insertion machinery (Sec) became overloaded upon CFTR production in L. lactis. Nutrients and ATP became limiting in the control cells as the culture entered the late exponential and stationary growth phases but this did not happen in the CFTR expressing cells, which had stopped growing upon induction. The different stress responses elicited in E. coli and L. lactis upon membrane protein production indicate that different strategies are needed to overcome low expression yields and toxicity.

  8. Coculture-inducible bacteriocin biosynthesis of different probiotic strains by dairy starter culture Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    Blaženka Kos

    2011-12-01

    Full Text Available Bacteriocins produced by probiotic strains effectively contribute to colonization ability of probiotic strains and facilitate their establishment in the competitive gut environment and also protect the gut from gastrointestinal pathogens. Moreover, bacteriocins have received considerable attention due to their potential application as biopreservatives, especially in dairy industry. Hence, the objective of this research was to investigate antimicrobial activity of probiotic strains Lactobacillus helveticus M92, Lactobacillus plantarum L4 and Enterococcus faecium L3, with special focus on their bacteriocinogenic activity directed towards representatives of the same or related bacterial species, and towards distant microorganisms including potential food contaminants or causative agents of gut infections. In order to induce bacteriocin production, probiotic cells were cocultivated with Lactococcus lactis subsp. lactis LMG 9450, one of the most important starter cultures in cheese production. The presence of bacteriocin coding genes was investigated by PCR amplification with sequence-specific primers for helveticin and was confirmed for probiotic strain L. helveticus M92. All examined probiotic strains have shown bacteriocinogenic activity against Staphylococcus aureus 3048, Staphylococcus aureus K-144, Escherichia coli 3014, Salmonella enterica serovar Typhimurium FP1, Bacillus subtilis ATCC 6633, Bacillus cereus TM2, which is an important functional treat of probiotic strains significant in competitive exclusion mechanism which provides selective advantage of probiotic strains against undesirable microorganisms in gastrointestinal tract of the host. According to obtained results, living cells of starter culture Lc. lactis subsp. lactis LMG 9450 induced bacteriocin production by examined probiotic strains but starter culture itself was not sensitive to bacteriocin activity.

  9. Development, molecular characterization and exploitation of the nisin controlled expression system in Lactococcus lactis.

    OpenAIRE

    Ruyter, de, D.J.

    1998-01-01

    Lactic acid bacteria are gram-positive bacteria that are widely used in a variety of dairy fermentation processes. Notably, strains of the lactic acid starter bacterium Lactococcus lactis are of great economic importance because of their world-wide use in cheese making. The characteristic aroma, flavor and texture of cheese develops during ripening of the cheese curd through the action of numerous enzymes derived from the cheese milk, the coagulant, and the starter and non-starter bacteria. R...

  10. Characterization of the Lactococcus lactis lactose genes and regulation of their expression.

    OpenAIRE

    Rooijen, van, J.

    1993-01-01

    An important trait of the lactic acid bacterium Lactococcus lactis , that is used in industrial dairy fermentations, is the conversion of lactose into lactic acid. The enzymatic steps involved in the breakdown of lactose, that is transported into the cell via a phosphoenolpyruvate-dependent lactose phosphotransferase system (PEP-PTS lac), have been well established (Fig. 1). However, except for the molecular cloning and characterization of the plasmid-located phospho-B-galactosidase gene (Boi...

  11. Milk fermentation by Lactococcus lactis with modified proteolytic systems to accumulate potentially bio-active peptides

    OpenAIRE

    Algaron, Florence; Miranda, Guy; Le Bars, Dominique; Monnet, Véronique

    2004-01-01

    International audience; The proteolytic system of lactic acid bacteria has been characterised in detail and numerous modified strains with null or increased specific proteolytic activities have been constructed or identified among natural strains. Based on this knowledge, our objective was to ferment milk with modified strains and produce mixtures of peptides with specific features corresponding to potential bio-activities. We used a collection of Lactococcus lactis negative mutants for pepti...

  12. Transcriptomic profile of aguR deletion mutant of Lactococcus lactis subsp. cremoris CECT 8666

    Directory of Open Access Journals (Sweden)

    Beatriz del Rio

    2015-12-01

    Full Text Available Lactococcus lactis subsp. cremoris CECT 8666 (formerly GE2-14 is a dairy strain that catabolizes agmatine (a decarboxylated derivative of arginine into the biogenic amine putrescine by the agmatine deiminase (AGDI pathway [1]. The AGDI cluster of L. lactis is composed by five genes aguR, aguB, aguD, aguA and aguC. The last four genes are responsible for the deamination of agmatine to putrescine and are co-transcribed as a single policistronic mRNA forming the catabolic operon aguBDAC [1]. aguR encodes a transmembrane protein that functions as a one-component signal transduction system that senses the agmatine concentration of the medium and accordingly regulates the transcription of aguBDAC [2], which is also transcriptionally regulated by carbon catabolic repression (CCR via glucose, but not by other sugars such as lactose and galactose [1,3]. Here we report the transcriptional profiling of the aguR gene deletion mutant (L. lactis subsp. cremoris CECT 8666 ∆aguR [2] compared to the wild type strain, both grown in M17 medium with galactose as carbon source and supplemented with agmatine. The transcriptional profiling data of AguR-regulated genes were deposited in the Gene Expression Omnibus (GEO database under accession no. GSE59514.

  13. Transcriptomic profile of aguR deletion mutant of Lactococcus lactis subsp. cremoris CECT 8666.

    Science.gov (United States)

    Del Rio, Beatriz; Linares, Daniel M; Redruello, Begoña; Martin, Maria Cruz; Fernandez, Maria; de Jong, Anne; Kuipers, Oscar P; Ladero, Victor; Alvarez, Miguel A

    2015-12-01

    Lactococcus lactis subsp. cremoris CECT 8666 (formerly GE2-14) is a dairy strain that catabolizes agmatine (a decarboxylated derivative of arginine) into the biogenic amine putrescine by the agmatine deiminase (AGDI) pathway [1]. The AGDI cluster of L. lactis is composed by five genes aguR, aguB, aguD, aguA and aguC. The last four genes are responsible for the deamination of agmatine to putrescine and are co-transcribed as a single policistronic mRNA forming the catabolic operon aguBDAC[1]. aguR encodes a transmembrane protein that functions as a one-component signal transduction system that senses the agmatine concentration of the medium and accordingly regulates the transcription of aguBDAC[2], which is also transcriptionally regulated by carbon catabolic repression (CCR) via glucose, but not by other sugars such as lactose and galactose [1], [3]. Here we report the transcriptional profiling of the aguR gene deletion mutant (L. lactis subsp. cremoris CECT 8666 ∆aguR) [2] compared to the wild type strain, both grown in M17 medium with galactose as carbon source and supplemented with agmatine. The transcriptional profiling data of AguR-regulated genes were deposited in the Gene Expression Omnibus (GEO) database under accession no. GSE59514.

  14. Use of Lactococcus lactis to enrich sourdough bread with γ-aminobutyric acid.

    Science.gov (United States)

    Bhanwar, Seema; Bamnia, Meenakshi; Ghosh, Moushumi; Ganguli, Abhijit

    2013-02-01

    Fried sourdough bread (bhatura) with an elevated amount of γ-aminobutyric acid (GABA) was produced using lactic acid bacteria (LAB). The LAB starter was screened and isolated from pickled yam showing highest GABA content and was identified as Lactococcus lactis subsp. lactis. The maximum GABA production in de Man Rogosa Sharpe (MRS) media supplemented with monosodium glutamate (MSG) was 110 mg/100 ml at pH 5, and 1-3% NaCl did not change the production of GABA significantly (p>0.05). When MSG was replaced with Vigna mungo in sourdough, the amount of GABA for bhatura was 226.22 mg/100 g representing about 10-fold increase. A sensory evaluation resulted as the overall general acceptability of bhatura to be 4.91 ± 0.03 on a five-point hedonic scale. Thus, the results indicated the potential of L. lactis as a LAB starter for the production of GABA-enriched bhatura. Although other physiological effects can be expected in the product, animal and clinical studies are mandatory prior to application of this food.

  15. The study of effect bacteriocin producing Lactoco ccus lactis on Listeria monocytogenes and Bacillus cereus

    Directory of Open Access Journals (Sweden)

    M. Mirhossieni, M.Sc

    2007-01-01

    Full Text Available AbstractBackground and purpose: Dairy products often associated with problems such as short shelf life and poor hygiene control. A novel approach is to utilize bacteriocin or bacteriocin producer strains, to control undesirable micro flora as Listeria monocytogenes and Bacillus cereus in foods. Hence, we studied the effect of nisin like producing Lactococcus lactis against Listeria monocytogenes and Bacillus cereus, in order to compare the isolated strain within different countries.Materials and Methods: In this research we studied the effect of nisin like producing Lactococcus lactis, with producer spot test method. We also used supernatant from 24 h culture of Lactoccus lactis. Moreover, we studied the effect of bacteriocin on Listeria monocytogenes and Bacillus cereus growth curves.Results: The growth of both strains was inhibited by the bacteriocin. Conclusion: According to our results, the bacteriocin could be used in liquid food with bacteriocin added directly or as a starter culture in fermentation. This would inhibit the growth of Listeria monocytogenes; furthermore, Bacillus cereus is used to reduce food poisoning for fermented food products.

  16. Enhance nisin yield via improving acid-tolerant capability of Lactococcus lactis F44

    Science.gov (United States)

    Zhang, Jian; Caiyin, Qinggele; Feng, Wenjing; Zhao, Xiuli; Qiao, Bin; Zhao, Guangrong; Qiao, Jianjun

    2016-01-01

    Traditionally, nisin was produced industrially by using Lactococcus lactis in the neutral fermentation process. However, nisin showed higher activity in the acidic environment. How to balance the pH value for bacterial normal growth and nisin activity might be the key problem. In this study, 17 acid-tolerant genes and 6 lactic acid synthetic genes were introduced in L. lactis F44, respectively. Comparing to the 2810 IU/mL nisin yield of the original strain F44, the nisin titer of the engineered strains over-expressing hdeAB, ldh and murG, increased to 3850, 3979 and 4377 IU/mL, respectively. These engineered strains showed more stable intracellular pH value during the fermentation process. Improvement of lactate production could partly provide the extra energy for the expression of acid tolerance genes during growth. Co-overexpression of hdeAB, murG, and ldh(Z) in strain F44 resulted in the nisin titer of 4913 IU/mL. The engineered strain (ABGL) could grow on plates with pH 4.2, comparing to the surviving pH 4.6 of strain F44. The fed-batch fermentation showed nisin titer of the co-expression L. lactis strain could reach 5563 IU/mL with lower pH condition and longer cultivation time. This work provides a novel strategy of constructing robust strains for use in industry process. PMID:27306587

  17. Heterologous protein secretion in Lactococcus lactis: a novel antigen delivery system

    Directory of Open Access Journals (Sweden)

    Langella P.

    1999-01-01

    Full Text Available Lactic acid bacteria (LAB are Gram-positive bacteria and are generally regarded as safe (GRAS organisms. Therefore, LAB could be used for heterologous protein secretion and they are good potential candidates as antigen delivery vehicles. To develop such live vaccines, a better control of protein secretion is required. We developed an efficient secretion system in the model LAB, Lactococcus lactis. Staphylococcal nuclease (Nuc was used as the reporter protein. We first observed that the quantity of secreted Nuc correlated with the copy number of the cloning vector. The nuc gene was cloned on a high-copy number cloning vector and no perturbation of the metabolism of the secreting strain was observed. Replacement of nuc native promoter by a strong lactococcal one led to a significant increase of nuc expression. Secretion efficiency (SE of Nuc in L. lactis was low, i.e., only 60% of the synthesized Nuc was secreted. Insertion of a synthetic propeptide between the signal peptide and the mature moiety of Nuc increased the SE of Nuc. On the basis of these results, we developed a secretion system and we applied it to the construction of an L. lactis strain which secretes a bovine coronavirus (BCV epitope-protein fusion (BCV-Nuc. BCV-Nuc was recognized by both anti-BCV and anti-Nuc antibodies. Secretion of this antigenic fusion is the first step towards the development of a novel antigen delivery system based on LAB-secreting strains.

  18. Biotechnological and safety characterization of Enterococcus lactis, a recently described species of dairy origin.

    Science.gov (United States)

    Morandi, Stefano; Silvetti, Tiziana; Brasca, Milena

    2013-01-01

    The biotechnological and safety properties of a recently described enterococcal species, Enterococcus lactis, were investigated. With regard to the technological properties, in milk all the strains tested had weak acidifying and proteolytic activities, generally medium reduction activity over 24 h (-102 mV detection did not identify any of the common genetic determinants for vancomycin, tetracycline and erythromycin resistance. The E. lactis strains showed good survival in simulated in vitro digestion and were able to inhibit the growth of Enterococcus durans, Enterococcus faecalis, Enterococcus faecium, Clostridium sporogenes, Clostridium tyrobutyricum and Pseudomonas syringae. Screening for enterocin structural genes showed that all isolates harboured the entP gene. The presence of nine virulence factor genes (cylA, asa1, gelE, hyl, esp, ace, efaA, hdc and tdc) was investigated by PCR and no virulence determinants were detected. This study highlights that the recently described E. lactis may be a potential source of novel strains with interesting features that could be used for fermented dairy foods.

  19. Purification and Characterization of Conjugated Bile Salt Hydrolase from Bifidobacterium longum BB536.

    Science.gov (United States)

    Grill, J; Schneider, F; Crociani, J; Ballongue, J

    1995-07-01

    Bifidobacterium species deconjugate taurocholic, taurodeoxycholic, taurochenodeoxycholic, glycocholic, glycodeoxycholic, and glycochenodeoxycholic acids. The enzyme level increases in the growth phase. No increase in activity is observed for the cytoplasmic enzyme after addition of conjugated bile acids to a stationary-phase culture. Conjugated bile salt hydrolase (BSH) was purified from Bifidobacterium longum BB536. Its apparent molecular mass in denaturing polyacrylamide gel electrophoresis was ca. 40,000 Da. The intact enzyme had a relative molecular weight of ca. 250,000 as determined by gel filtration chromatography, suggesting that the native BSH of B. longum is probably a hexamer. The purified enzyme is active towards both glycine and taurine conjugates of cholate, deoxycholate, and chenodeoxycholate. The pH optimum is in the range of 5.5 to 6.5. A loss of BSH activity is observed after incubation at temperatures higher than 42(deg)C; at 60(deg)C, 50% of the BSH activity is lost. The importance of free sulfhydryl groups at the enzyme active center is suggested. For B. longum BB536, no significant difference in the initial rate of deconjugation and enzymatic efficiency appears between bile salts. The enzymatic efficiency is higher for B. longum BB536 than for other genera. In this paper, a new method which permits a display of BSH activity directly on polyacrylamide gels is described; this method confirms the molecular weight obtained for B. longum BB536 BSH.

  20. In vitro kinetic analysis of oligofructose consumption by Bacteroides and Bifidobacterium spp. indicates different degradation mechanisms.

    Science.gov (United States)

    Van der Meulen, Roel; Makras, Lefteris; Verbrugghe, Kristof; Adriany, Tom; De Vuyst, Luc

    2006-02-01

    The growth of pure cultures of Bacteroides thetaiotaomicron LMG 11262 and Bacteroides fragilis LMG 10263 on fructose and oligofructose was examined and compared to that of Bifidobacterium longum BB536 through in vitro laboratory fermentations. Gas chromatography (GC) analysis was used to determine the different fractions of oligofructose and their degradation during the fermentation process. Both B. thetaiotaomicron LMG 11262 and B. fragilis LMG 10263 were able to grow on oligofructose as fast as on fructose, succinic acid being the major metabolite produced by both strains. B. longum BB536 grew slower on oligofructose than on fructose. Acetic acid and lactic acid were the main metabolites produced when fructose was used as the sole energy source. Increased amounts of formic acid and ethanol were produced when oligofructose was used as an energy source at the cost of lactic acid. Detailed kinetic analysis revealed a preferential metabolism of the short oligofructose fractions (e.g., F2 and F3) for B. longum BB536. After depletion of the short fractions, the larger oligofructose fractions (e.g., F4, GF4, F5, GF5, and F6) were metabolized, too. Both Bacteroides strains did not display such a preferential metabolism and degraded all oligofructose fractions simultaneously, transiently increasing the fructose concentration in the medium. This suggests a different mechanism for oligofructose breakdown between the strain of Bifidobacterium and both strains of Bacteroides, which helps to explain the bifidogenic nature of inulin-type fructans.

  1. Creation of the Probiotic Consortium on the Base of Strains of Bifidobacterium spp.

    Directory of Open Access Journals (Sweden)

    Kozhakhmetov, S. S.

    2009-01-01

    Full Text Available Nowadays, a widespread circulation of disbiotic conditions among the population of all ages in Kazakhstan requires an active development in industry for both preparations and products with probiotic properties. The gained bacterial isolates, Bifidobacterium adolescentis 180, B. breve 204, B. breve 584 and B. breve 587 were used in our researches and screening showed they possess high probiotic properties. The consortium possesses strong antimicrobial activity to pathogenic and potentially-pathogenic microflora, insulated during disbacteriosis, as well as from vagina and urea. They are able to produce vitamin B12 and also have antimutagenic activity. As a result, the consortium on the base of strains of Bifidobacterium spp. was received, possessing the following advantages: contains live mass of microbial, antagonistically active strains B. breve and B. adolescentis; contains more than 10^9 alive Bifidobacteria; does not contain plasmids, which means that it could not be a carrier of antibiotic stability for Gram-positive receptive pathogenic and potentially-pathogenic microflora.

  2. Effects of cigarette smoke condensate on the production and characterization of exopolysaccharides by Bifidobacterium

    Directory of Open Access Journals (Sweden)

    Jinqiang Hu

    2015-06-01

    Full Text Available The aim of the study was to investigate the effect of cigarette smoke on the production and characterization of exopolysaccharides (EPSs produced by Bifidobacterium. Cigarettes of Shanhua brand (nicotine: 1.1 mg, tar: 11 mg were utilized to prepare a cigarette smoke condensate (CSC. The standard strain of Bifidobacterium animalis was cultured in MRS media under anaerobic addition of CSC. The results showed that CSC significantly decreased the growth of B. animalis as well as EPSs and acetic acid production. Furthermore, two EPSs fractions (Fr-I and Fr-II were isolated and purified for chemical and molecular determination. By comparison with control, CSC was found to be of great impact on EPSs carbohydrate composition. The molecular weight mass of Fr-I changed from 3.33×105 g/mol (without CSC to 2.99×105 (with CSC. In conclusion, in vitro studies revealed that CSC was directly able to affect the production of metabolites for B. animalis, which could be an essential factor in certain pathological disorders.

  3. Comparative genomic analysis of 45 type strains of the genus Bifidobacterium: a snapshot of its genetic diversity and evolution.

    Directory of Open Access Journals (Sweden)

    Zhihong Sun

    Full Text Available Bifidobacteria are well known for their human health-promoting effects and are therefore widely applied in the food industry. Members of the Bifidobacterium genus were first identified from the human gastrointestinal tract and were then found to be widely distributed across various ecological niches. Although the genetic diversity of Bifidobacterium has been determined based on several marker genes or a few genomes, the global diversity and evolution scenario for the entire genus remain unresolved. The present study comparatively analyzed the genomes of 45 type strains. We built a robust genealogy for Bifidobacterium based on 402 core genes and defined its root according to the phylogeny of the tree of bacteria. Our results support that all human isolates are of younger lineages, and although species isolated from bees dominate the more ancient lineages, the bee was not necessarily the original host for bifidobacteria. Moreover, the species isolated from different hosts are enriched with specific gene sets, suggesting host-specific adaptation. Notably, bee-specific genes are strongly associated with respiratory metabolism and are potential in helping those bacteria adapt to the oxygen-rich gut environment in bees. This study provides a snapshot of the genetic diversity and evolution of Bifidobacterium, paving the way for future studies on the taxonomy and functional genomics of the genus.

  4. In vitro study of the prebiotic xylooligosaccharide (XOS) on the growth of Bifidobacterium spp and Lactobacillus spp.

    Science.gov (United States)

    Li, Zhaoping; Summanen, Paula H; Komoriya, Tomoe; Finegold, Sydney M

    2015-01-01

    We recently demonstrated that XOS increased the counts of Bifidobacterium in vivo without increasing Lactobacillus in healthy adults. In the current study, we evaluated the effect of XOS on the growth of 35 Bifidobacterium and 29 Lactobacillus strains in in vitro conditions. Bacteria were identified by 16S rRNA sequence analysis. The growth stimulation was determined by agar dilution technique on plates containing two-fold serial dilutions of XOS (100-0.1 mg/ml). The growth of 86% of Bifidobacterium strains was stimulated at 1.56 mg/ml XOS and 100% at 6.25 mg/ml XOS. The growth of 38% of Lactobacillus strains was stimulated at 1.56 mg/ml XOS and 62% at 6.25 mg/ml XOS; 31% of Lactobacillus were not stimulated by XOS. Our results further suggest that XOS may be beneficial in stimulating intestinal Bifidobacterium without having much effect on Lactobacillus. The potential role for XOS in managing obesity should be investigated further.

  5. Post-sensitization administration of non-digestible oligosaccharides and Bifidobacterium breve M-16V reduces allergic symptoms in mice

    NARCIS (Netherlands)

    van Esch, Betty C A M; Abbring, Suzanne; Diks, Mara A P; Dingjan, Gemma M; Harthoorn, Lucien F; Vos, A Paul; Garssen, Johan

    2016-01-01

    To support dietary management of severe cow's milk allergic infants, a synbiotic mixture of non-digestible oligosaccharides and Bifidobacterium breve M-16V (B. breve) was designed from source materials that are completely cow's milk-free. It was investigated whether this specific synbiotic concept c

  6. Non-Fusion and Fusion Expression of β-Galactosidase from Lactobacillus bulgaricus in Lactococcus lactis

    Institute of Scientific and Technical Information of China (English)

    CHUAN WANG; CHAO-WU ZHANG; HENG-CHUAN LIU; QIAN YU; XIAO-FANG PEI

    2008-01-01

    Objective To construct four recombinant Lactococcus lactis strains exhibiting high β-galactosidase activity in fusion or non-fusion ways, and to study the influence factors for their protein expression and secretion. Methods The gene fragments encoding β-galactosidase from two strains of Lactobacillus bulgaricus, wch9901 isolated from yogurt and 1.1480 purchased from the Chinese Academy of Sciences, were amplified and inserted into lactococcal expression vector pMG36e. For fusion expression, the open reading frame of the β-galactosidase gene was amplified, while for non-fusion expression, the open reading frame of the β-galactosidase gene was amplified with its native Shine-Dalgarno sequence upstream. The start codon of the β-galactosidase gene partially overlapped with the stop codon of vector origin open reading frame. Then, the recombinant plasmids were transformed into Escherichia coli DH5α and Lactococcus lactis subsp, lactis MG1363 and confirmed by determining β-galactosidase activities. Results The non-fusion expression plasmids showed a significantly higher β-galactosidase activity in transformed strains than the fusion expression plasmids. The highest enzyme activity was observed in Lactococcus lactis transformed with the non-fusion expression plasmids which were inserted into the β-galactosidase gene from Lactobacillus bulgaricus wch9901. The β-galactosidase activity was 2.75 times as high as that of the native counterpart. In addition, β-galactosidase expressed by recombinant plasmids in Lactococcus lactis could be secreted into the culture medium. The highest secretion rate (27.1%) was observed when the culture medium contained 20 g/L of lactose. Conclusion Different properties of the native bacteria may have some effects on the protein expression of recombinant plasmids. Non-fusion expression shows a higher enzyme activity in host bacteria. There may be a ost-related weak secretion signal peptide gene within the structure gene of Lb

  7. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of a health claim related to a combination of Lactobacillus helveticus CNCM I-1722, Bifidobacterium longum subsp. infantis CNCM I-3424, Bifidobacterium bifidum CNCM I-3426 and fructo

    DEFF Research Database (Denmark)

    Tetens, Inge

    of Lactobacillus helveticus CNCM I-1722, Bifidobacterium longum subsp. infantis CNCM I-3424, Bifidobacterium bifidum CNCM I-3426 and fructo-oligosaccharides from sucrose and immune defence against pathogens. The food constituent that is the subject of the health claim, a combination of L. helveticus CNCM I-1722, B...

  8. Lactococcus lactis NCC 2287 Alleviates Food Allergic Manifestations in Sensitized Mice by Reducing IL-13 Expression Specifically in the Ileum

    Directory of Open Access Journals (Sweden)

    Adrian W. Zuercher

    2012-01-01

    Full Text Available Objective. Utilizing a food allergy murine model, we have investigated the intrinsic antiallergic potential of the Lactococcus lactis NCC 2287 strain. Methods. BALB/c mice were sensitized at weekly intervals with ovalbumin (OVA plus cholera toxin (CT by the oral route for 7 weeks. In this model, an oral challenge with a high dose of OVA at the end of the sensitization period leads to clinical symptoms. Lactococcus lactis NCC 2287 was given to mice via the drinking water during sensitization (prevention phase or after sensitization (management phase. Results. Lactococcus lactis NCC 2287 administration to sensitized mice strikingly reduced allergic manifestations in the management phase upon challenge, when compared to control mice. No preventive effect was observed with the strain. Lactococcus lactis NCC 2287 significantly decreased relative expression levels of the Th-2 cytokine, IL-13, and associated chemokines CCL11 (eotaxin-1 and CCL17 (TARC in the ileum. No effect was observed in the jejunum. Conclusion/Significance. These results taken together designate Lactococcus lactis NCC 2287 as a candidate probiotic strain appropriate in the management of allergic symptoms.

  9. Display of the Viral Epitopes on Lactococcus lactis: A Model for Food Grade Vaccine against EV71.

    Science.gov (United States)

    Varma, Nadimpalli Ravi S; Toosa, Haryanti; Foo, Hooi Ling; Alitheen, Noorjahan Banu Mohamed; Nor Shamsudin, Mariana; Arbab, Ali S; Yusoff, Khatijah; Abdul Rahim, Raha

    2013-01-01

    In this study, we have developed a system for display of antigens of Enterovirus type 71 (EV71) on the cell surface of L. lactis. The viral capsid protein (VP1) gene from a local viral isolate was utilized as the candidate vaccine for the development of oral live vaccines against EV71 using L. lactis as a carrier. We expressed fusion proteins in E. coli and purified fusion proteins were incubated with L. lactis. We confirmed that mice orally fed with L. lactis displaying these fusion proteins on its surface were able to mount an immune response against the epitopes of EV71. This is the first example of an EV71 antigen displayed on the surface of a food grade organism and opens a new perspective for alternative vaccine strategies against the EV71. We believe that the method of protein docking utilized in this study will allow for more flexible presentations of short peptides and proteins on the surface of L. lactis to be useful as a delivery vehicle.

  10. Display of the Viral Epitopes on Lactococcus lactis: A Model for Food Grade Vaccine against EV71

    Directory of Open Access Journals (Sweden)

    Nadimpalli Ravi S. Varma

    2013-01-01

    Full Text Available In this study, we have developed a system for display of antigens of Enterovirus type 71 (EV71 on the cell surface of L. lactis. The viral capsid protein (VP1 gene from a local viral isolate was utilized as the candidate vaccine for the development of oral live vaccines against EV71 using L. lactis as a carrier. We expressed fusion proteins in E. coli and purified fusion proteins were incubated with L. lactis. We confirmed that mice orally fed with L. lactis displaying these fusion proteins on its surface were able to mount an immune response against the epitopes of EV71. This is the first example of an EV71 antigen displayed on the surface of a food grade organism and opens a new perspective for alternative vaccine strategies against the EV71. We believe that the method of protein docking utilized in this study will allow for more flexible presentations of short peptides and proteins on the surface of L. lactis to be useful as a delivery vehicle.

  11. Lactococcus lactis NCC 2287 Alleviates Food Allergic Manifestations in Sensitized Mice by Reducing IL-13 Expression Specifically in the Ileum

    Science.gov (United States)

    Zuercher, Adrian W.; Weiss, Marietta; Holvoet, Sébastien; Moser, Mireille; Moussu, Hélène; van Overtvelt, Laurence; Horiot, Stéphane; Moingeon, Philippe; Nutten, Sophie; Prioult, Guénolée; Singh, Anurag; Mercenier, Annick

    2012-01-01

    Objective. Utilizing a food allergy murine model, we have investigated the intrinsic antiallergic potential of the Lactococcus lactis NCC 2287 strain. Methods. BALB/c mice were sensitized at weekly intervals with ovalbumin (OVA) plus cholera toxin (CT) by the oral route for 7 weeks. In this model, an oral challenge with a high dose of OVA at the end of the sensitization period leads to clinical symptoms. Lactococcus lactis NCC 2287 was given to mice via the drinking water during sensitization (prevention phase) or after sensitization (management phase). Results. Lactococcus lactis NCC 2287 administration to sensitized mice strikingly reduced allergic manifestations in the management phase upon challenge, when compared to control mice. No preventive effect was observed with the strain. Lactococcus lactis NCC 2287 significantly decreased relative expression levels of the Th-2 cytokine, IL-13, and associated chemokines CCL11 (eotaxin-1) and CCL17 (TARC) in the ileum. No effect was observed in the jejunum. Conclusion/Significance. These results taken together designate Lactococcus lactis NCC 2287 as a candidate probiotic strain appropriate in the management of allergic symptoms. PMID:21961022

  12. Protective vaccination against infectious bursal disease virus with whole recombinant Kluyveromyces lactis yeast expressing the viral VP2 subunit.

    Directory of Open Access Journals (Sweden)

    Marina Arnold

    Full Text Available Here we report on vaccination approaches against infectious bursal disease (IBD of poultry that were performed with complete yeast of the species Kluyveromyces lactis (K. lactis. Employing a genetic system that enables the rapid production of stably transfected recombinant K. lactis, we generated yeast strains that expressed defined quantities of the virus capsid forming protein VP2 of infectious bursal disease virus (IBDV. Both, subcutaneous as well as oral vaccination regiments with the heat-inactivated but otherwise untreated yeast induced IBDV-neutralizing antibodies in mice and chickens. A full protection against a subsequent IBDV infection was achieved by subcutaneous inoculation of only milligram amounts of yeast per chicken. Oral vaccination also generated protection: while mortality was observed in control animals after virus challenge, none of the vaccinees died and ca. one-tenth were protected as indicated by the absence of lesions in the bursa of Fabricius. Recombinant K. lactis was thus indicated as a potent tool for the induction of a protective immune response by different applications. Subcutaneously applied K. lactis that expresses the IBDV VP2 was shown to function as an efficacious anti-IBD subunit vaccine.

  13. AguR is a transmembrane transcription activator of the putrescine biosynthesis operon in Lactococcus lactis, and acts in response to agmatine concentration

    NARCIS (Netherlands)

    Linares, Daniel M; Del Rio, Beatriz; Redruello, Begoña; Ladero, Victor; Martin, Ma Cruz; de Jong, Anne; Kuipers, Oscar P; Fernandez, Maria; Alvarez, Miguel A

    2015-01-01

    Dairy industry fermentative processes mostly use Lactococcus lactis as a starter. However, some dairy L. lactis strains produce putrescine - a biogenic amine that raises food safety and spoilage concerns - via the agmatine deiminase pathway (AGDI). The enzymatic activities responsible for putrescine

  14. Transcriptional activation of the glycolytic las operon and catabolite repression of the gal operon in Lactococcus lactis are mediated by the catabolite control protein CcpA

    NARCIS (Netherlands)

    Luesink, Evert J.; Herpen, René E.M.A. van; Grossiord, Benoît P.; Kuipers, Oscar P.; Vos, Willem M. de

    1998-01-01

    The Lactococcus lactis ccpA gene, encoding the global regulatory protein CcpA, was identified and characterized. Northern blot and primer extension analyses showed that the L. lactis ccpA gene is constitutively transcribed from a promoter that does not contain a cre sequence. Inactivation of the ccp

  15. Identification and functional characterization of the Lactococcus lactis CodY-regulated branched-chain amino acid permease BcaP (CtrA)

    NARCIS (Netherlands)

    den Hengst, CD; Groeneveld, M; Kuipers, OP; Kok, J; Hengst, Chris D. den

    2006-01-01

    Transcriptome analyses have previously revealed that a gene encoding the putative amino acid transporter CtrA (YhdG) is one of the major targets of the pleiotropic regulator CodY in Lactococcus lactis and Bacillus subtilis. The role of ctrA in L. lactis was further investigated with respect to both

  16. Antigen-presenting cells exposed to Lactobacillus acidophilus NCFM, Bifidobacterium bifidum BI-98, and BI-504 reduce regulatory T cell activity

    DEFF Research Database (Denmark)

    Schmidt, Esben Gjerløff Wedebye; Claesson, Mogens Helweg; Jensen, Simon Skjøde

    2010-01-01

    BACKGROUND:: The effect in vitro of six different probiotic strains including Lactobacillus acidophilus NCFM, Lactobacillus salivarius Ls-33, Lactobacillus paracasei subsp. paracasei YS8866441, Lactobacillus plantarum Lp-115, Bifidobacterium bifidum BI-504 and BI-98 was studied on splenic...

  17. The effect of soymilk intake on the fecal microbiota, particularly Bifidobacterium species, and intestinal environment of healthy adults: a pilot study.

    Science.gov (United States)

    Fujisawa, Tomohiko; Ohashi, Yuji; Shin, Ryoichi; Narai-Kanayama, Asako; Nakagaki, Takenori

    2017-01-01

    The influence of soymilk on the fecal microbiota, particularly Bifidobacterium species, and metabolic activities were investigated in eight healthy adult humans. During the soymilk intake period, the number of bifidobacteria in feces was significantly higher (penvironment.

  18. Effects of Bifidobacterium longum BB536 on lipid profile and histopathological changes in hypercholesterolaemic rats.

    Science.gov (United States)

    Al-Sheraji, S H; Amin, I; Azlan, A; Manap, M Y; Hassan, F A

    2015-01-01

    The present study investigated the effects of Bifidobacterium longum BB536 on lipid profile, liver and kidney function, and body fat in hypercholesterolaemic rats. 40 Sprague-Dawley rats were randomly divided into five groups. The negative control group received a standard diet. The positive control group received a cholesterol-enriched diet, whereas the intervention groups received a cholesterol-enriched diet supplemented with B. longum BB536 alone or in combination with inulin or Mangifera pajang fibrous polysaccharides. After 8 weeks, plasma lipids, and liver and kidney function were tested. Intake of the cholesterol-enriched diet increased total cholesterol, alanine aminotransferase, gamma-glutamyl transferase, creatinine, urea, liver weight, adipose tissue weight, liver lipid deposition and adipocyte size. B. longum BB536 supplementation significantly reduced total cholesterol, liver lipid deposition and adipocyte size, and positively affected liver and kidney function. These effects were significantly increased in the presence of inulin and M. pajang fibrous polysaccharides.

  19. Characterization and Application of BiLA, a Psychrophilic α-Amylase from Bifidobacterium longum.

    Science.gov (United States)

    Lee, Hye-Won; Jeon, Hye-Yeon; Choi, Hye-Jeong; Kim, Na-Ri; Choung, Woo-Jae; Koo, Ye-Seul; Ko, Dam-Seul; You, SangGuan; Shim, Jae-Hoon

    2016-04-06

    In this study, a novel α-amylase was cloned from Bifidobacterium longum and named BiLA. The enzyme exhibited optimal activity at 20 °C and a pH value of 5.0. Kinetic analysis using various carbohydrate substrates revealed that BiLA had the highest k(cat/)K(m) value for amylose. Interestingly, analysis of the enzymatic reaction products demonstrated that BiLA specifically catalyzed the hydrolysis of oligosaccharides and starches up to G5 from the nonreducing ends. To determine whether BiLA can be used to generate slowly digestible starch (SDS), starch was treated with BiLA, and the kinetic parameters were analyzed using porcine pancreatic α-amylase (PPA) and amyloglucosidase (AMG). Compared to normal starch, BiLA-treated starch showed lower k(cat)/K(m) values with PPA and AMG, suggesting that BiLA is a potential candidate for the production of SDS.

  20. Optimization of nitrogen source for Bifidobacterium bifidum using response surface methodology

    Directory of Open Access Journals (Sweden)

    Chen He

    2016-06-01

    Full Text Available In order to improve the viable counts of Bifidobacterium bifidum BB01 in the liquid medium, the Central Composite Design (CCD was used to optimize the nitrogen source in the medium of B. bifidum BB01. The results showed that the nitrogen source composition of B. bifidum BB01 was: peptone 0.9%, yeast extracts 0.3%, beef paste 0.7%. Under the optimal conditions, the viable counts of B. bifidum BB01 reached (2.49±0.06×109CFU/mL after cultured at 18h, which was 42.97% higher than MRS (lactose, and 12.85% higher than the optimized MRS medium (carbon source and prebiotics were optimized. Therefore, the CCD used in this study is workable for promoting the growth of B. bifidum BB01.

  1. Genetic analyses of the antibiotic resistance of Bifidobacterium bifidum strain Yakult YIT 4007.

    Science.gov (United States)

    Sato, Takashi; Iino, Tohru

    2010-02-28

    Bifidobacterium bifidum strain Yakult YIT 4007 (abbreviated as B. bifidum YIT 4007) is a commercial strain and resistant to erythromycin, neomycin, and streptomycin. Resistances to these antibiotics were endowed by sequential isolation of resistant mutants from its susceptible progenitor strain YIT 4001. Comparison of nucleotide sequences of various candidate genes of both strains led us to find that B. bifidum YIT 4007 had mutations on three copies of 23S ribosomal RNA genes, an 8 bp deletion of the rluD gene for pseudouridine synthase, and a mutation on the rpsL gene for ribosomal protein S12. The responsibility of these mutations to antibiotic resistances was supported by analyses of newly isolated mutants resistant to these antibiotics. The antibiotic resistances of B. bifidum YIT 4007 were evidently acquired by mutations of the structural genes on the chromosome and not associated with mobile genetic elements like insertion sequences, phages, and plasmids.

  2. Applications of microencapsulated Bifidobacterium longum with Eleutherine americana in fresh milk tofu and pineapple juice.

    Science.gov (United States)

    Phoem, Atchara N; Chanthachum, Suphitchaya; Voravuthikunchai, Supayang P

    2015-04-01

    Bifidobacterium longum was microencapsulated by extrusion technique and added in fresh milk tofu and pineapple juice. Microencapsulation of B. longum with Eleutherine americana extract, oligosaccharides extract, and commercial fructo-oligosaccharides was assessed for the bacterial survival after sequential exposure to simulated gastric and intestinal juices, and refrigeration storage. Microencapsulated B. longum with the extract and oligosaccharides extract in the food products showed better survival than free cells under adverse conditions. Sensory analysis demonstrated that the products containing co-encapsulated bacterial cells were more acceptable by consumers than free cells. Pineapple juice prepared with co-encapsulated cells had lower values for over acidification, compared with the juice with free cells added. This work suggested that microencapsulated B. longum with E. americana could enhance functional properties of fresh milk tofu and pineapple juice.

  3. Applications of Microencapsulated Bifidobacterium Longum with Eleutherine Americana in Fresh Milk Tofu and Pineapple Juice

    Directory of Open Access Journals (Sweden)

    Atchara N. Phoem

    2015-04-01

    Full Text Available Bifidobacterium longum was microencapsulated by extrusion technique and added in fresh milk tofu and pineapple juice. Microencapsulation of B. longum with Eleutherine americana extract, oligosaccharides extract, and commercial fructo-oligosaccharides was assessed for the bacterial survival after sequential exposure to simulated gastric and intestinal juices, and refrigeration storage. Microencapsulated B. longum with the extract and oligosaccharides extract in the food products showed better survival than free cells under adverse conditions. Sensory analysis demonstrated that the products containing co-encapsulated bacterial cells were more acceptable by consumers than free cells. Pineapple juice prepared with co-encapsulated cells had lower values for over acidification, compared with the juice with free cells added. This work suggested that microencapsulated B. longum with E. americana could enhance functional properties of fresh milk tofu and pineapple juice.

  4. CRECIMIENTO, SOBREVIVENCIA Y ADAPTACIÓN DE Bifidobacterium infantis A CONDICIONES ÁCIDAS

    OpenAIRE

    L. Mayorga-Reyes; P. Bustamante-Camilo; A. Gutiérrez-Nava; E. Barranco-Florido; A. Azaola-Espinosa

    2009-01-01

    La acidez es una condición ambiental comúnmente encontrada por las bacterias presentes en productos lácteos fermentados y el tracto gastrointestinal. En este estudio, las células de Bifidobacterium infantis de 24 h de fermentación se inocularon en medios de cultivo con pH iniciales de 7.0, 4.0, 3.0 y 2.0 durante 24 h. Conforme el pH inicial disminuye, la población celular activa disminuyó hasta tres órdenes de magnitud. A pH 4.0 las células se mostraron estables durante las primeras 5 h de fe...

  5. Optimization of Fermentation Conditions for Nisin Production by Lactococcus lactis N302%Nisin生产菌株Lactococcus lactis N302的发酵优化

    Institute of Scientific and Technical Information of China (English)

    李瑞青; 轩辕铮铮; 姜德洲; 苏俊杰; 徐海津; 张秀明; 乔明强

    2011-01-01

    对一株Nisin生产菌株Lactococcus lactis N302现有培养基进行了氮源替代,并采用Plackett-Burman(PB)法和中心复合设计(Central Composite Design)对影响其发酵生产Nisin的6个培养条件进行筛选优化.PB实验表明,蔗糖、初始pH值和酵母粉是影响Nisin效价的三个关键因素.对三因素进行中心复合设计,经响应面法优化分析(RSM)确定了L.Lactis N302发酵生产Nisin的最优条件为:蔗糖13.7g.L-1,初始pH值7.74,酵母粉25.7g.L-1,大豆蛋白胨10.0g.L-1,K2HPO410.0g.L-1,接种量3%.优化后Nisin效价较优化前提高了7.2%.小试(10 L)研究表明,分批发酵18h、补碱分批发酵16h菌株L.lactis N302单位Nisin效价最高,分别为4 597.03 IU.mL-1和8 773.34 IU.mL-1.%Nisin is a bacteriocin widely used in food industry as an effective food preservative. High nisin production was aimed by optimizing the fermentation conditions of Lactococcus lactis N302. First, soybean peptone was used the main nitrogen source of the culture medium instead of peptone. Then, the Plackett-Burman design (PB) and the path of steepest ascent method were applied to investigate the main factors that affect the yield of nisin, and to find the optimum region of the response. The results indicated that sucrose, initial pH value and yeast extract were the significant factors for nisin production. Central composite experimental design and response surface methodology (RSM) were further adopted to derive a statistical model for optimizing the fermentation conditions. The optimum fermentation conditions were found to be sucrose 13. 7 g · L-1, initial pH value 7. 74, yeast extract 25. 7 g · L-1, soybean peptone 10 g · L-1, K2HPO410 g · L-1, inoculum size 3%. The nisin yield increased by 7. 2% compared to the no-optimized conditions. Finally, 10 liter batch and pH fed-batch fermentation with the optimized conditions were carried out. The maximum nisin yield was achieved at 18 h for batch fermentation and 16 h for fed

  6. Genetically engineered Lactococcus lactis protect against house dust mite allergy in a BALB/c mouse model.

    Directory of Open Access Journals (Sweden)

    Chunqing Ai

    Full Text Available Mucosal vaccine based on lactic acid bacteria is an attractive concept for the prevention and treatment of allergic diseases, but their mechanisms of action in vivo are poorly understood. Therefore, we sought to investigate how recombinant major dust mite allergen Der p2-expressing Lactococcus lactis as a mucosal vaccine induced the immune tolerance against house dust mite allergy in a mouse model.Three strains of recombinant L. lactis producing Der p2 in different cell components (extracellular, intracellular and cell wall were firstly constructed. Their prophylactic potential was evaluated in a Der p2-sensitised mouse model, and immunomodulation properties at the cellular level were determined by measuring cytokine production in vitro.Der p2 expressed in the different recombinant L. lactis strains was recognized by a polyclonal anti-Der p2 antibody. Oral treatment with the recombinant L. lactis prior sensitization significantly prevented the development of airway inflammation in the Der p2-sensitized mice, as determined by the attenuation of inflammatory cells infiltration in the lung tissues and decrease of Th2 cytokines IL-4 and IL-5 levels in bronchoalveolar lavage. In addition, the serum allergen-specific IgE levels were significantly reduced, and the levels of IL-4 in the spleen and mesenteric lymph nodes cell cultures were also markedly decreased upon allergen stimulation in the mice fed with the recombinant L. lactis strains. These protective effects correlated with a significant up-regulation of regulatory T cells in the mesenteric lymph nodes.Oral pretreatment with live recombinant L. lactis prevented the development of allergen-induced airway inflammation primarily by the induction of specific mucosal immune tolerance.

  7. Characterization of the Fermentation Process and the Inhibition Effect of Lactobacillus lactis in Staphylococcus aureus and Staphylococcus epidermidis

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    Henry Jurado-Gámez

    2015-09-01

    Full Text Available The fermentative process and in vitro inhibition of L. lactis in Staphylococcus aureus and Staphylococcus epidermidis were assessed. The growth of L. lactis at three pH (2.5, 4.5 and 7, bile salts (0.5, 1 and 2 %, bovine bile (1 and 1.2 % and two temperatures (38 and 45 °C were evaluated. Peptides and organic acids in supernatant of L. lactis by HPLC were determined. Fermentation kinetics was carried out, evaluating: pH, total sugar, protein and lactic acid. An antibiogram of dicloxacilin, cefepime, penilicin and cefalotin was made. The inhibition of L. lactis and its supernatant were defined in pathogenic strains. The best growth was at a pH of 2.5 (3 × 1012 UFC/ml; of 1 × 1010 and 4 × 109 UFC/ml for 0.5 % of bile salts and 1.2 % of bovine bile, respectively; of 3.5 × 1013 and 3.4 × 1013 UFC/ml for 38 and 45 °C, respectively. The HPLC determined the peptides VAR-TIR-VAR and lactic acid (83.11 %. The fermentation kinetics determined the exponential phase at 14:24 h with a value of 77 × 1010 UFC/ ml, pH values of 4.284, 2.33 mg/ml sugar, 1.44 mg/ml protein and acidity of 0.79 %. It was found that S. aureus and S. epidermidis were sensitive to all antibiotics. The pathogenic bacteria were resistant to the lactic strain, but S. epidermidis was sensitive to the supernatant of L. lactis. The conclusion is that Lactobacillus lactis showed adequate growth capacity, good fermentation parameters and inhibitory effect in strains of S. aureus and S. epidermidis in in vitro conditions.

  8. Improved adhesive properties of recombinant bifidobacteria expressing the Bifidobacterium bifidum-specific lipoprotein BopA

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    Gleinser Marita

    2012-06-01

    Full Text Available Abstract Background Bifidobacteria belong to one of the predominant bacterial groups in the intestinal microbiota of infants and adults. Several beneficial effects on the health status of their human hosts have been demonstrated making bifidobacteria interesting candidates for probiotic applications. Adhesion of probiotics to the intestinal epithelium is discussed as a prerequisite for colonisation of and persistence in the gastrointestinal tract. Results In the present study, 15 different strains of bifidobacteria were tested for adhesion. B. bifidum was identified as the species showing highest adhesion to all tested intestinal epithelial cell (IEC lines. Adhesion of B. bifidum S17 to IECs was strongly reduced after treatment of bacteria with pronase. These results strongly indicate that a proteinaceous cell surface component mediates adhesion of B. bifidum S17 to IECs. In silico analysis of the currently accessible Bifidobacterium genomes identified bopA encoding a lipoprotein as a B. bifidum-specific gene previously shown to function as an adhesin of B. bifidum MIMBb75. The in silico results were confirmed by Southern Blot analysis. Furthermore, Northern Blot analysis demonstrated that bopA is expressed in all B. bifidum strains tested under conditions used to cultivate bacteria for adhesion assays. The BopA gene was successfully expressed in E. coli and purified by Ni-NTA affinity chromatography as a C-terminal His6-fusion. Purified BopA had an inhibitory effect on adhesion of B. bifidum S17 to IECs. Moreover, bopA was successfully expressed in B. bifidum S17 and B. longum/infantis E18. Strains overexpressing bopA showed enhanced adhesion to IECs, clearly demonstrating a role of BopA in adhesion of B. bifidum strains. Conclusions BopA was identified as a B. bifidum-specific protein involved in adhesion to IECs. Bifidobacterium strains expressing bopA show enhanced adhesion. Our results represent the first report on recombinant

  9. Intraspecies Genomic Diversity and Long-Term Persistence of Bifidobacterium longum.

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    Andrei V Chaplin

    Full Text Available Members of genus Bifidobacterium are Gram-positive bacteria, representing a large part of the human infant microbiota and moderately common in adults. However, our knowledge about their diversity, intraspecific phylogeny and long-term persistence in humans is still limited. Bifidobacterium longum is generally considered to be the most common and prevalent species in the intestinal microbiota. In this work we studied whole genome sequences of 28 strains of B. longum, including 8 sequences described in this paper. Part of these strains were isolated from healthy children during a long observation period (up to 10 years between isolation from the same patient. The three known subspecies (longum, infantis and suis could be clearly divided using sequence-based phylogenetic methods, gene content and the average nucleotide identity. The profiles of glycoside hydrolase genes reflected the different ecological specializations of these three subspecies. The high impact of horizontal gene transfer on genomic diversity was observed, which is possibly due to a large number of prophages and rapidly spreading plasmids. The pan-genome characteristics of the subspecies longum corresponded to the open pan-genome model. While the major part of the strain-specific genetic loci represented transposons and phage-derived regions, a large number of cell envelope synthesis genes were also observed within this category, representing high variability of cell surface molecules. We observed the cases of isolation of high genetically similar strains of B. longum from the same patients after long periods of time, however, we didn't succeed in the isolation of genetically identical bacteria: a fact, reflecting the high plasticity of microbiota in children.

  10. Intraspecies Genomic Diversity and Long-Term Persistence of Bifidobacterium longum

    Science.gov (United States)

    Chaplin, Andrei V.; Efimov, Boris A.; Smeianov, Vladimir V.; Kafarskaia, Lyudmila I.; Pikina, Alla P.; Shkoporov, Andrei N.

    2015-01-01

    Members of genus Bifidobacterium are Gram-positive bacteria, representing a large part of the human infant microbiota and moderately common in adults. However, our knowledge about their diversity, intraspecific phylogeny and long-term persistence in humans is still limited. Bifidobacterium longum is generally considered to be the most common and prevalent species in the intestinal microbiota. In this work we studied whole genome sequences of 28 strains of B. longum, including 8 sequences described in this paper. Part of these strains were isolated from healthy children during a long observation period (up to 10 years between isolation from the same patient). The three known subspecies (longum, infantis and suis) could be clearly divided using sequence-based phylogenetic methods, gene content and the average nucleotide identity. The profiles of glycoside hydrolase genes reflected the different ecological specializations of these three subspecies. The high impact of horizontal gene transfer on genomic diversity was observed, which is possibly due to a large number of prophages and rapidly spreading plasmids. The pan-genome characteristics of the subspecies longum corresponded to the open pan-genome model. While the major part of the strain-specific genetic loci represented transposons and phage-derived regions, a large number of cell envelope synthesis genes were also observed within this category, representing high variability of cell surface molecules. We observed the cases of isolation of high genetically similar strains of B. longum from the same patients after long periods of time, however, we didn’t succeed in the isolation of genetically identical bacteria: a fact, reflecting the high plasticity of microbiota in children. PMID:26275230

  11. Licheniocin 50.2 and Bacteriocins from Lactococcus lactis subsp. lactis biovar. diacetylactis BGBU1-4 Inhibit Biofilms of Coagulase Negative Staphylococci and Listeria monocytogenes Clinical Isolates

    Science.gov (United States)

    Draganic, Veselin; Lozo, Jelena; Beric, Tanja; Kojic, Milan; Arsic, Biljana; Garalejic, Eliana; Djukic, Slobodanka; Stankovic, Slavisa

    2016-01-01

    Background Coagulase negative staphylococci (CoNS) and Listeria monocytogenes have important roles in pathogenesis of various genital tract infections and fatal foetomaternal infections, respectively. The aim of our study was to investigate the inhibitory effects of two novel bacteriocins on biofilms of CoNS and L. monocytogenes genital isolates. Methods The effects of licheniocin 50.2 from Bacillus licheniformis VPS50.2 and crude extract of bacteriocins produced by Lactococcus lactis subsp. lactis biovar. diacetylactis BGBU1-4 (BGBU1-4 crude extract) were evaluated on biofilm formation and formed biofilms of eight CoNS (four S. epidermidis, two S. hominis, one S. lugdunensis and one S. haemolyticus) and 12 L. monocytogenes genital isolates. Results Licheniocin 50.2 and BGBU1-4 crude extract inhibited the growth of both CoNS and L. monocytogenes isolates, with MIC values in the range between 200–400 AU/ml for licheniocin 50.2 and 400–3200 AU/ml for BGBU1-4 crude extract. Subinhibitory concentrations (1/2 × and 1/4 × MIC) of licheniocin 50.2 inhibited biofilm formation by all CoNS isolates (p bacteriocins in concentrations of 100 AU/mL and 200 AU/mL reduced the amount of 24 h old CoNS and L. monocytogenes biofilms (p bacteriocins have potential to be used for genital application, to prevent biofilm formation and/or to eradicate formed biofilms, and consequently reduce genital and neonatal infections by CoNS and L. monocytogenes. PMID:27930711

  12. Cloning and verification of the Lactococcus lactis pyrG gene and characterization of the gene product, CTP synthase

    DEFF Research Database (Denmark)

    Wadskov-Hansen, Steen Lyders Lerche; Willemoës, M.; Martinussen, Jan

    2001-01-01

    The pyrG gene of Lactococcus lactis subsp. cremoris, encoding CTP synthase, has been cloned and sequenced. It is flanked upstream by an open reading frame showing homology to several aminotransferases and downstream by an open reading frame of unknown function. L. lactis strains harboring disrupted...... of a functional cdd gene encoding cytidine deaminase. A characterization of the enzyme revealed similar properties as found for CTP synthases from other organisms. However, unlike the majority of CTP synthases the lactococcal enzyme can convert dUTP to dCTP, although a half saturation concentration of 0.6 m...

  13. Glucose metabolism in Lactococcus lactis MG1363 under different aeration conditions: Requirement of acetate to sustain growth under microaerobic conditions

    DEFF Research Database (Denmark)

    Nordkvist, Mikkel; Jensen, N.B.S.; Villadsen, John

    2003-01-01

    Lactococcus lactis subsp. lactis MG1363 was grown in batch cultures on a defined medium with glucose as the energy source under different aeration conditions, namely, anaerobic conditions, aerobic conditions, and microaerobic conditions with a dissolved oxygen tension of 5% (when saturation...... with air was used as the reference). The maximum specific growth rate was high (0.78 to 0.91 h(-1)) under all aeration conditions but decreased with increasing aeration, and more than 90% of the glucose was converted to lactate. However, a shift in by-product formation was observed. Increasing aeration...

  14. Transcriptome profiling of Lactococcus lactis subsp. cremoris CECT 8666 in response to agmatine

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    Beatriz del Rio

    2016-03-01

    Full Text Available The dairy strain Lactococcus lactis subsp. cremoris CECT 8666 (formerly GE2-14 synthesizes the biogenic amine putrescine from agmatine via the agmatine deiminase (AGDI pathway [1]. The AGDI cluster of L. lactis is composed by five genes aguR, aguB, aguD, aguA and aguC. The last four genes are co-transcribed as a single policistronic mRNA forming the catabolic operon aguBDAC, which encodes the proteins necessary for agmatine uptake and its conversion into putrescine [1,2]. The first gene of the cluster, aguR, encodes a transmembrane protein that functions as a one-component signal transduction system that senses the agmatine concentration of the medium and accordingly regulates the transcription of aguBDAC [2]. The catabolic operon aguBDAC is transcriptionally activated by agmatine [2] and transcriptionally regulated by carbon catabolite repression (CCR via glucose, but not by other sugars such as lactose or galactose [1,3]. On the contrary, the transcription of the aguR regulatory gene is not subject to CCR regulation [1,3] nor is regulated by agmatine [2]. In this study we report the transcriptional profiling of L. lactis subsp. cremoris CECT 8666 grown in M17 medium with galactose (GalM17 as carbon source and supplemented with agmatine, compared to that of the strain grown in the same culture medium without agmatine. The transcriptional profiling data of agmatine-regulated genes were deposited in the Gene Expression Omnibus (GEO database under Accession no. GSE74808.

  15. Transcriptome profiling of Lactococcus lactis subsp. cremoris CECT 8666 in response to agmatine.

    Science.gov (United States)

    Del Rio, Beatriz; Redruello, Begoña; Martin, M Cruz; Fernandez, Maria; de Jong, Anne; Kuipers, Oscar P; Ladero, Victor; Alvarez, Miguel A

    2016-03-01

    The dairy strain Lactococcus lactis subsp. cremoris CECT 8666 (formerly GE2-14) synthesizes the biogenic amine putrescine from agmatine via the agmatine deiminase (AGDI) pathway [1]. The AGDI cluster of L. lactis is composed by five genes aguR, aguB, aguD, aguA and aguC. The last four genes are co-transcribed as a single policistronic mRNA forming the catabolic operon aguBDAC, which encodes the proteins necessary for agmatine uptake and its conversion into putrescine [1], [2]. The first gene of the cluster, aguR, encodes a transmembrane protein that functions as a one-component signal transduction system that senses the agmatine concentration of the medium and accordingly regulates the transcription of aguBDAC[2]. The catabolic operon aguBDAC is transcriptionally activated by agmatine [2] and transcriptionally regulated by carbon catabolite repression (CCR) via glucose, but not by other sugars such as lactose or galactose [1], [3]. On the contrary, the transcription of the aguR regulatory gene is not subject to CCR regulation [1], [3] nor is regulated by agmatine [2]. In this study we report the transcriptional profiling of L. lactis subsp. cremoris CECT 8666 grown in M17 medium with galactose (GalM17) as carbon source and supplemented with agmatine, compared to that of the strain grown in the same culture medium without agmatine. The transcriptional profiling data of agmatine-regulated genes were deposited in the Gene Expression Omnibus (GEO) database under Accession no. GSE74808.

  16. Quantitative comparison of transient growth of Saccharomyces cerevisiae, Saccharomyces kluyveri, and Kluyveromyces lactis.

    Science.gov (United States)

    Herwig, Christoph; Von Stockar, Urs

    2003-03-30

    A multitude of metabolic regulations occur in yeast, particularly under dynamic process conditions, such as under sudden glucose excess. However, quantification of regulations and classification of yeast strains under these conditions have yet to be elucidated, which requires high-frequency and consistent quantification of the metabolic response. The present study aimed at quantifying the dynamic regulation of the central metabolism of strains Saccharomyces cerevisiae, S. kluyveri, and Kluyveromyces lactis upon sudden glucose excess, accomplished by a shift-up in dilution rate inside of the oxidative region using a small metabolic flux model. It was found that, under transient growth conditions, S. kluyveri behaved like K. lactis, while classification using steady-state conditions would position S. kluyveri close to S. cerevisiae. For transient conditions and based on the observation whether excess glucose is initially used for catabolism (energy) or anabolism (carbon), we propose to classify strains into energy-driven, such as S. cerevisiae, and carbon-driven, such as S. kluyveri and K. lactis, strains. Furthermore, it was found that the delayed onset of fermentative catabolism in carbon-driven strains is a consequence of low catabolic flux and the initial shunt of glucose in non-nitrogen-containing biomass constituents. The MFA model suggests that energy limitation forced the cell to ultimately increase catabolic flux, while the capacity of oxidative catabolism is not sufficient to process this flux oxidatively. The combination of transient experiments and its exploitation with reconciled intrinsic rates using a small metabolic model could corroborate earlier findings of metabolic regulations, such as tight glucose control in carbon-driven strains and transient changes in biomass composition, as well as explore new regulations, such as assimilation of ethanol before glucose. The benefit from using small metabolic flux models is the richness of information and the

  17. Nisin inducible production of listeriolysin O in Lactococcus lactis NZ9000

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    Griffin Brendan T

    2008-07-01

    Full Text Available Abstract Background Listeria monocytogenes is a well-characterized food-borne pathogen that infects pregnant women and immunocompromised individuals. Listeriolysin O (LLO is the major virulence factor of the pathogen and is often used as a diagnostic marker for detection of L. monocytogenes. In addition, LLO represents a potent antigen driving T cell-mediated immunity during infection. In the present work, Lactococcus lactis NZ9000 was used as an expression host to hyper-produce LLO under inducible conditions using the NICE (NIsin Controlled Expression system. We created a modified pNZ8048 vector encoding a six-His-tagged LLO downstream of the strong inducible PnisA promoter. Results The constructed vector (pNZPnisA:CYTO-LLO was expressed in L. lactis NZ9000 and was best induced at mid-log phase with 0.2% v/v nisin for 4 h statically at 30°C. Purification of the His-tagged LLO was accomplished by Ni-NTA affinity chromatography and functionality was confirmed through haemolytic assays. Total LLO yield (measured as total protein content was 4.43–5.9 mg per litre culture and the haemolytic activity was still detectable after 8 months of storage at 4°C. Conclusion The LLO production method described in this work provides an approach to efficient LLO production in the Gram-positive Lactococcus bacterium to yield a significant source of the protein for research and diagnostic applications. Expression of LLO in L. lactis has a number of benefits over E. coli which may facilitate both in vivo and in vitro applications of this system.

  18. Enhancement of nisin production by Lactococcus lactis in periodically re-alkalized cultures.

    Science.gov (United States)

    Guerra, Nelson Pérez; Castro, Lorenzo Pastrana

    2003-10-01

    Synthesis of nisin as well as biomass production by Lactococcus lactis subsp. lactis CECT (Colección Española de Cultivos Tipo) 539 on both hydrolysed mussel-processing waste and whey medium were followed in three fixed volume fed-batch fermentations, with re-alkalization cycles. The two cultures on mussel-processing waste (MPW) were fed with a 240 g/l concentrated glucose and with a concentrated MPW (about 100 g of glucose/l). The culture on whey was fed with a mixture of concentrated whey (48 g of total sugars/l) and a 400 g/l concentrated lactose. The three cultures were mainly characterized with higher nisin titres [49.7, 109.6 and 124.7 bacteriocin activity units (AU)/ml respectively] compared with the batch process on de Man, Rogosa and Sharpe [(1960) J. Appl. Bacteriol. 23, 130-135] medium (49.6 AU/ml), MPW (9.5 AU/ml) and whey (22.5 AU/ml) [1 AU/ml is the amount of antibacterial compound needed to obtain 50% growth inhibition (LD50) compared with control tubes]. In the three fed-batch cultures a shift from homolactic to mixed-acid fermentation was observed, and other products (acetic acid, butane-2,3-diol or ethanol) in addition to lactic acid were detectable in the medium. However, their contributions to the total antibacterial activity of the post-incubates (the cell-free culture supernatant obtained at the end of the fermentation process) of L. lactis CECT 539 against Carnobacterium piscicola CECT 4020 were very low.

  19. Enzymatic synthesis and characterization of hydroquinone galactoside using Kluyveromyces lactis lactase.

    Science.gov (United States)

    Kim, Go-Eun; Lee, Jin-Ha; Jung, Sun-Hwa; Seo, Eun-Seong; Jin, Sheng-De; Kim, Ghahyun J; Cha, Jaeho; Kim, Eui-Joong; Park, Ki-Deok; Kim, Doman

    2010-09-08

    Hydroquinone galactoside (HQ-Gal) as a potential skin whitening agent was synthesized by the reaction of lactase (beta-galactosidase) from Kluyveromyces lactis, Aspergillus oryzae, Bacillus circulans, and Thermus sp. with lactose as a donor and HQ as an acceptor. Among these lactases, the acceptor reaction involving HQ and lactose with K. lactis lactase showed a higher conversion ratio to HQ-Gal (60.27%). HQ-Gal was purified using butanol partitioning and silica gel column chromatography. The structure of the purified HQ-Gal was determined by nuclear magnetic resonance, and the ionic product was observed at m/z 295 (C12H16O7Na)+ using matrix assisted laser desorption ionization time-of-flight mass spectrometry. HQ-Gal was identified as 4-hydroxyphenyl-beta-d-galactopyranoside. The optimum conditions for HQ-Gal synthesis by K. lactis determined using response surface methodology were 50 mM HQ, 60 mM lactose, and 20 U mL(-1) lactase. These conditions produced a yield of 2.01 g L(-1) HQ-Gal. The half maximal inhibitory concentration (IC50) of diphenylpicrylhydrazyl scavenging activity was 3.31 mM, indicating a similar antioxidant activity compared to beta-arbutin (IC50=3.95 mM). The Ki value of HQ-Gal (0.75 mM) against tyrosinase was smaller than that of beta-arbutin (Ki=1.97 mM), indicating its superiority as an inhibitor. HQ-Gal inhibited (23%) melanin synthesis without being significantly toxic to the cells, while beta-arbutin exhibited only 8% reduction of melanin synthesis in B16 melanoma cells compared with the control. These results indicate that HQ-Gal may be a suitable functional component in the cosmetics industry.

  20. Role of Bifidobacterium bifidum and plant food extracts in improving microflora and biochemical and cytogenetic parameters in adjuvant arthritis.

    Directory of Open Access Journals (Sweden)

    Donya, S. M.

    2011-09-01

    Full Text Available The aim of the present research was to discover plant food extracts and probiotics that may have bioactivity towards chronic inflammation. Three plant food extract mixtures expected to be rich in phenolic compounds, carotenoids and tocopherols were prepared. The anti-inflammatory activity of the different mixtures as well as probiotic bacteria (Bifidobacterium bifidum were evaluated in adjuvant arthritis in rats. The anti-inflammatory effect, mechanism of action and safety of the three mixtures and Bifidobacterium bifidum were studied by measuring the size of inflammation and the determination of inflammatory and oxidative stress biomarkers, colonic bacteria profile and specific cytogenetic parameters. The contents of tocopherols, β-carotene and phenolic compounds in the mixtures were determined. The results show that the tested mixtures and Bifidobacterium bifidum possess promising anti-inflammatory effects. The mechanism of action seems to involve a reduction in oxidative stress and inflammatory biomarkers and an effect on colonic microflora. Genotoxicity and DNA fragmentation induced by adjuvant arthritis were prevented after supplementation with the tested mixtures.El objetivo de la presente investigación fue encontrar extractos de alimentos vegetales y probióticos que puedan tener bioactividad hacía la inflamación crónica. Mezclas de tres extractos de alimentos vegetales conocidos por su riqueza en compuestos fenólicos, carotenoides y tocoferoles han sido preparadas. La actividad anti-inflamatoria de las diferentes mezclas y de bacterias probióticas (Bifidobacterium bifidum fue evaluada en artritis adyuvante en ratas. El efecto anti-inflamatorio, mecanismo de acción y salubridad de las tres mezclas y de Bifidobacterium bifidum ha sido estudiado mediante la medida del tamaño de la inflamación y la determinación de biomarcadores de inflamación y estrés oxidativo, del perfil de bacterias del colón y de parámetros citogen

  1. Phosphoglycerate Mutase Is a Highly Efficient Enzyme without Flux Control in Lactococcus lactis

    DEFF Research Database (Denmark)

    Solem, Christian; Petranovic, D.; Købmann, Brian

    2010-01-01

    The glycolytic enzyme phosphoglycerate mutase (PGM), which catalyzes the conversion of 3-phosphoglycerate to 2-phosphoglycerate, was examined in Lactococcus lactis with respect to its function, kinetics and glycolytic flux control. A library of strains with PGM activities ranging between 15......-465% of the wild-type level was constructed by replacing the native promoter of pgm with synthetic promoters of varying strengths. The specific growth rate and glucose flux were found to be maximal at the wild-type level at which PGM had no flux control. Low flux control of PGM was found on mixed acid fluxes...

  2. Mechanism of flavin reduction in the class 1A dihydroorotate dehydrogenase from Lactococcus lactis

    DEFF Research Database (Denmark)

    Fagan, Rebecca L; Jensen, Kaj Frank; Björnberg, Olof;

    2007-01-01

    is concerted or stepwise was addressed for the class 1A enzyme from Lactococcus lactis by determining kinetic isotope effects (KIEs) on flavin reduction in anaerobic stopped-flow experiments. Isotope effects were determined at two pH values. At pH 7.0, KIEs were approximately 2-fold for DHO labeled singly...... mutants was extremely slow compared to that of the wild type; the rate of reduction increased with pH, showing no sign of a plateau. Interestingly, double-deuterium isotope effects on the Cys130Ser mutant also showed a concerted mechanism for flavin reduction....

  3. Cloning and Characterization of upp, a Gene Encoding Uracil Phosphoribosyltransferase from Lactococcus lactis

    DEFF Research Database (Denmark)

    Martinussen, Jan; Hammer, Karin

    1994-01-01

    Uracil phosphoribosyltransferase catalyzes the key reaction in the salvage of uracil in many microorganisms. The gene encoding uracil phosphoribosyltransferase (upp) was cloned from Lactococcus lactis subsp. cremoris MG1363 by complementation of an Escherichia coli mutant. The gene was sequenced...... construction of an internal deletion, a upp mutant was constructed by a double-crossover event. This implicated the utilization of a plasmid with a thermosensitive origin of replication and a new and easy way to screen for double crossover events in both gram-positive and gram-negative bacterial strains...

  4. Cloning and Expression of the Lactococcus lactis purDEK Genes, Required for Growth in Milk

    DEFF Research Database (Denmark)

    Nilsson, Dan; Kilstrup, Mogens

    1998-01-01

    An operon containing the genes purD and purE and part of the purK gene was cloned from the facultative anaerobic gram positive bacterium Lactococcus lactis by complementation of the purD mutation in Escherichia coli SO609. The genes encode enzymes in the de novo pathway of purine nucleotides....... The expression of the genes was regulated approximately 35-fold at the transcription level by the availability of purines in the growth medium. Deletion analysis of the nucleotide region upstream of purD indicated that a region of 145 bp is enough to give regulated expression of the reporter lacLM genes, which...

  5. Metabolic behavior of Lactococcus lactis MG1363 in microaerobic continuous cultivation at a low dilution rate

    DEFF Research Database (Denmark)

    Jensen, Niels B.S.; Melchiorsen, Claus Rix; Jochumsen, Kirsten Væver;

    2001-01-01

    Minute amounts of oxygen were supplied to a continuous cultivation of Lactococcus lactis subsp. cremoris MG1363 grown on a defined glucose-limited medium at a dilution rate of 0.1 h(-1). More than 80% of the carbon supplied with glucose ended up in fermentation products other than lactate. Addition...... dioxide and acetate to replace formate and ethanol as catabolic end products while hardly affecting the production of either acetoin or lactate. The negative impact of oxygen on the synthesis of pyruvate formate lyase was confirmed. Moreover, oxygen was shown to down regulate the protein level of alcohol...

  6. Fine tuning of the lactate and diacetyl production through promoter engineering in Lactococcus lactis.

    Directory of Open Access Journals (Sweden)

    Tingting Guo

    Full Text Available Lactococcus lactis is a well-studied bacterium widely used in dairy fermentation and capable of producing metabolites with organoleptic and nutritional characteristics. For fine tuning of the distribution of glycolytic flux at the pyruvate branch from lactate to diacetyl and balancing the production of the two metabolites under aerobic conditions, a constitutive promoter library was constructed by randomizing the promoter sequence of the H(2O-forming NADH oxidase gene in L. lactis. The library consisted of 30 promoters covering a wide range of activities from 7,000 to 380,000 relative fluorescence units using a green fluorescent protein as reporter. Eleven typical promoters of the library were selected for the constitutive expression of the H(2O-forming NADH oxidase gene in L. lactis, and the NADH oxidase activity increased from 9.43 to 58.17-fold of the wild-type strain in small steps of activity change under aerobic conditions. Meanwhile, the lactate yield decreased from 21.15 ± 0.08 mM to 9.94 ± 0.07 mM, and the corresponding diacetyl production increased from 1.07 ± 0.03 mM to 4.16 ± 0.06 mM with the intracellular NADH/NAD(+ ratios varying from 0.711 ± 0.005 to 0.383 ± 0.003. The results indicated that the reduced pyruvate to lactate flux was rerouted to the diacetyl with an almost linear flux variation via altered NADH/NAD(+ ratios. Therefore, we provided a novel strategy to precisely control the pyruvate distribution for fine tuning of the lactate and diacetyl production through promoter engineering in L. lactis. Interestingly, the increased H(2O-forming NADH oxidase activity led to 76.95% lower H(2O(2 concentration in the recombinant strain than that of the wild-type strain after 24 h of aerated cultivation. The viable cells were significantly elevated by four orders of magnitude within 28 days of storage at 4°C, suggesting that the increased enzyme activity could eliminate H(2O(2 accumulation and prolong cell survival.

  7. THE APOPTOSIS OF EXPERIMENTAL COLORECTAL CARCINOMA CELLS INDUCED BY PEPTIDOGLYCAN OF BIFIDOBACTERIUM AND THE EXPRESSION OF APOPTOTIC REGULATING GENES

    Institute of Scientific and Technical Information of China (English)

    WANG Li-sheng; PAN Ling-jia; SHI Li; SUN Yong; ZHANG Ya-li; ZHOU Dian-yuan

    1999-01-01

    Objective: To explore the antitumor mechanisms of whole peptidoglycan of bifidobacterium. Methods: The apoptotic cells and the positive expression of bcl-2 and bax oncoprotein were studied nude mice transplantation tumors of colorectal carcinoma by employing in situ end labeling technique and immunohistochemical staining. Results:The apoptotic cell density, the positive rate and the staining intensity of bax oncoprotein of the transplantation tumor of colorectal carcinoma in the whole peptidoglycan injection group were significantly higher when compared with the tumor control group. The positive rate of bcl-2 oncoprotein in the whole peptidoglycan injection group was obviously lower than that in the tumor control group (P<0.01).Conclusion: Whole peptidoglycan of Bifidobacterium bifidum could induce cell apoptosis of nude mice transplantation tumors of colorectal carcinoma by downregulating the expression of the bcl-2 gene and upregulating the expression of the bax gene.

  8. Survival of Bifidobacterium longum and its effect on physicochemical properties and sensorial attributes of white brined cheese.

    Science.gov (United States)

    Gursoy, Oguz; Gokce, Ramazan; Con, Ahmet Hilmi; Kinik, Ozer

    2014-11-01

    Survival of the probiotic adjunct culture Bifidobacterium longum and cheese starters during ripening of white brined cheese, effect of the probiotic culture on physicochemical properties and sensorial attributes of cheeses were investigated throughout 90 d of ripening. Bifidobacterium longum were able to survive at higher levels (>10(7 )cfu/g cheese) than the therapeutic minimum (10(6)-10(7 )cfu/g cheese) after 90 d and did not have any negative effect on the survival of Streptococcus spp. (including common cheese starters). Incorporation of the probiotic adjunct into white brined cheese and high levels of their survival rates during ripening had an insignificant effect on the composition of cheeses. Results indicated that white brined cheese is a suitable food matrix for the delivery of B. longum used in this study, and white brined cheeses with B. longum may be considered as a probiotic dairy product.

  9. Bifidobacterium breve reduces apoptotic epithelial cell shedding in an exopolysaccharide and MyD88-dependent manner

    Science.gov (United States)

    Hughes, K. R.; Harnisch, L. C.; Alcon-Giner, C.; Mitra, S.; Wright, C. J.; Ketskemety, J.

    2017-01-01

    Certain members of the microbiota genus Bifidobacterium are known to positively influence host well-being. Importantly, reduced bifidobacterial levels are associated with inflammatory bowel disease (IBD) patients, who also have impaired epithelial barrier function, including elevated rates of apoptotic extrusion of small intestinal epithelial cells (IECs) from villi—a process termed ‘cell shedding’. Using a mouse model of pathological cell shedding, we show that mice receiving Bifidobacterium breve UCC2003 exhibit significantly reduced rates of small IEC shedding. Bifidobacterial-induced protection appears to be mediated by a specific bifidobacterial surface exopolysaccharide and interactions with host MyD88 resulting in downregulation of intrinsic and extrinsic apoptotic responses to protect epithelial cells under highly inflammatory conditions. Our results reveal an important and previously undescribed role for B. breve, in positively modulating epithelial cell shedding outcomes via bacterial- and host-dependent factors, supporting the notion that manipulation of the microbiota affects intestinal disease outcomes. PMID:28123052

  10. Effect of repeated oral administration of Bifidobacterium longum BB536 on apomorphine-induced rearing behavior in mice

    OpenAIRE

    ORIKASA, Shuzo; NABESHIMA, Kazumi; Iwabuchi, Noriyuki; Xiao, Jin-zhong

    2016-01-01

    Schizophrenia is a chronic psychiatric illness. Disruption of the dopaminergic system has been suggested to be the pathogenic cause of this disease. The effect of Bifidobacterium longum BB536 (BB536) on schizophrenic behavior was investigated in an animal model. Daily administration of BB536 (109 CFU/mouse, p.o. for 2 weeks) was found to reduce rearing behavior augmented by the dopamine receptor agonist apomorphine and to decrease the resting level of plasma corticosterone and the ratio of ky...

  11. Randomized controlled trial of live lactobacillus acidophilus plus bifidobacterium bifidum in prophylaxis of diarrhea during radiotherapy in cervical cancer patients

    OpenAIRE

    Chitapanarux, Imjai; Chitapanarux, Taned; Traisathit, Patrinee; Kudumpee, Sudkaneung; Tharavichitkul, Ekkasit; Lorvidhaya, Vicharn

    2010-01-01

    Background Radiation-induced diarrhea is frequently observed during pelvic radiotherapy. This study was performed to determine the ability of a probiotic containing live lactobacillus acidophilus plus bifidobacterium bifidum to reduce the incidence of radiation-induced diarrhea in locally advanced cervical cancer patients. Methods Patients who were undergoing pelvic radiotherapy concurrent with weekly cisplatin were randomly assigned to a study drug or placebo, in a double-blind study. Diarrh...

  12. Chimeras of mature pediocin PA-1 fused to the signal peptide of enterocin P permits the cloning, production, and expression of pediocin PA-1 in Lactococcus lactis.

    Science.gov (United States)

    Martín, María; Gutiérrez, Jorge; Criado, Raquel; Herranz, Carmen; Cintas, Luis M; Hernández, Pablo E

    2007-12-01

    Chimeras of pediocin PA-1 (PedA-1), a bacteriocin produced by Pediococcus acidilactici PLBH9, fused to the signal peptide of enterocin P (EntP), a sec-dependent bacteriocin produced by Enterococcus faecium P13, permitted the production of PedA-1 in Lactococcus lactis. Chimeric genes encoding the EntP signal peptide (SP(entP)) fused to mature PedA-1 (pedA), with or without its immunity gene (pedB), were cloned into the expression vector pMG36c to generate the recombinant plasmids pMPP9 (SP(entP):pedA) and pMPP14i (SP(entP):pedA + pedB). Transformation of competent L. lactis subsp. lactis IL1403, L. lactis subsp. cremoris NZ9000, and L. lactis subsp. lactis DPC5598 with the recombinant plasmids has permitted the detection and quantitation of PedA-1 and the coproduction of nisin A and PedA-1 in supernatants of producer cells with specific anti-PedA-1 antibodies and a noncompetitive indirect enzyme-linked immunosorbent assay. Recombinant L. lactis hosts carrying pMPP9 or pMPP14i displayed antimicrobial activity, suggesting that mature PedA-1 fused to SP(EntP) is the minimum requirement for the synthesis, processing, and secretion of biologically active PedA-1 in L. lactis. However, the production and antimicrobial activity of the PedA-1 produced by L. lactis was lower than that produced by the P. acidilactici control strains.

  13. Molecular Characterization of a Recombinant Manganese Superoxide Dismutase from Lactococcus lactis M4

    Directory of Open Access Journals (Sweden)

    Boon Hooi Tan

    2014-01-01

    Full Text Available A superoxide dismutase (SOD gene of Lactococcus lactis M4 was cloned and expressed in a prokaryotic system. Sequence analysis revealed an open reading frame of 621 bp which codes for 206 amino acid residues. Expression of sodA under T7 promoter exhibited a specific activity of 4967 U/mg when induced with 1 mM of isopropyl-β-D-thiogalactopyranoside. The recombinant SOD was purified to homogeneity by immobilised metal affinity chromatography and Superose 12 gel filtration chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot analyses of the recombinant SOD detected a molecular mass of approximately 27 kDa. However, the SOD was in dimer form as revealed by gel filtration chromatography. The purified recombinant enzyme had a pI of 4.5 and exhibited maximal activity at 25°C and pH 7.2. It was stable up to 45°C. The insensitivity of this lactococcal SOD to cyanide and hydrogen peroxide established that it was a MnSOD. Although it has 98% homology to SOD of L. lactis IL1403, this is the first elucidated structure of lactococcal SOD revealing active sites containing the catalytic manganese coordinated by four ligands (H-27, H-82, D-168, and H-172.

  14. Single-Step Partial Purification of Intracellular β-Galactosidase from Kluyveromyces lactis Using Microemulsion Droplets.

    Science.gov (United States)

    Mazı, Bekir G; Hamamcı, Haluk; Ogrydziak, David M; Dungan, Stephanie R

    2016-11-01

    Partial purification of β-galactosidase from the crude extract of Kluyveromyces lactis was carried out using water-in-isooctane microemulsions formed by the anionic surfactant, sodium di-ethylhexyl sulfosuccinate (Aerosol OT). In order to obtain the crude extract, yeast cells of K. lactis were disrupted by a cell disrupter and separated. The purification of β-galactosidase from the extract by a recently developed one-step reversed micellar (i.e., microemulsion-based) extraction method was then tested, by measuring total protein mass and enzyme activity in the product stream and by analyzing its composition using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography. Effects of salt concentration, protein concentration, and pH on the extraction were investigated. Using this approach, a 5.4-fold purification of β-galactosidase was achieved with 96 % total activity recovery, using a feed containing crude extract and 50 mM K-phosphate buffer (pH 7.5) and 50 mM KCl. Gel filtration chromatography showed that the single extraction was successful at removing low molecular weight impurity proteins (molecular weight (MW) extract.

  15. Isolation and characterization of exopolysaccharide from Leuconostoc lactis KC117496 isolated from idli batter.

    Science.gov (United States)

    Saravanan, Chinnashanmugam; Shetty, Prathap Kumar H

    2016-09-01

    Diverse exopolysaccharide (EPS)-producing isolates were isolated from an Indian acidic fermented food (Idli) based on the colony morphology. One of the EPS-producing microflora (Leuconostoc lactis KC117496) was selected for further characterization using FT-IR, HPTLC, AFM, SEM, TGA and XRD analysis. FT-IR spectroscopy revealed the α-d-glucose nature of the EPS. HPTLC analysis confirmed the presence of only glucose monomers, indicating the glucan nature of EPS. NMR spectra revealed the presence of 95% α-(1→6) and 5% branching α-(1→3) linkages. The SEM and AFM showed smooth surfaces and compact structure. TGA results showed higher degradation temperature of 272.01°C. XRD analysis proved the 33.4% crystalline nature of the EPS. Water solubility index and water-holding capacity of EPS are 14.2±0.208% and 117±7.5%. All the above characteristics of the EPS produced by L. lactis showed that the EPS is of a good-quality polysaccharide with potential applications in the food industry.

  16. Interaction of benzoate pyrimidine analogues with class 1A dihydroorotate dehydrogenase from Lactococcus lactis

    DEFF Research Database (Denmark)

    Wolfe, Abigail E; Thymark, Majbritt; Gattis, Samuel G;

    2007-01-01

    Dihydroorotate dehydrogenases (DHODs) catalyze the oxidation of dihydroorotate to orotate in the only redox reaction in pyrimidine biosynthesis. The pyrimidine binding sites are very similar in all structurally characterized DHODs, suggesting that the prospects for identifying a class-specific in......Dihydroorotate dehydrogenases (DHODs) catalyze the oxidation of dihydroorotate to orotate in the only redox reaction in pyrimidine biosynthesis. The pyrimidine binding sites are very similar in all structurally characterized DHODs, suggesting that the prospects for identifying a class......-specific inhibitor directed against this site are poor. Nonetheless, two compounds that bind specifically to the Class 1A DHOD from Lactococcus lactis, 3,4-dihydroxybenzoate (3,4-diOHB) and 3,5-dihydroxybenzoate (3,5-diOHB), have been identified [Palfey et al. (2001) J. Med. Chem. 44, 2861-2864]. The mechanism...... of inhibitor binding to the Class 1A DHOD from L. lactis has now been studied in detail and is reported here. Titrations showed that 3,4-diOHB binds more tightly at higher pH, whereas the opposite is true for 3,5-diOHB. Isothermal titration calorimetry and absorbance spectroscopy showed that 3,4-diOHB ionizes...

  17. Time-resolved genetic responses of Lactococcus lactis to a dairy environment.

    Science.gov (United States)

    Bachmann, Herwig; de Wilt, Leonie; Kleerebezem, Michiel; van Hylckama Vlieg, Johan E T

    2010-05-01

    Lactococcus lactis is one of main bacterial species found in mixed dairy starter cultures for the production of semi-hard cheese. Despite the appreciation that mixed cultures are essential for the eventual properties of the manufactured cheese the vast majority of studies on L. lactis were carried out in laboratory media with a pure culture. In this study we applied an advanced recombinant in vivo expression technology (R-IVET) assay in combination with a high-throughput cheese-manufacturing protocol for the identification and subsequent validation of promoter sequences specifically induced during the manufacturing and ripening of cheese. The system allowed gene expression measurements in an undisturbed product environment without the use of antibiotics and in combination with a mixed strain starter culture. The utilization of bacterial luciferase as reporter enabled the real-time monitoring of gene expression in cheese for up to 200 h after the cheese-manufacturing process was initiated. The results revealed a number of genes that were clearly induced in cheese such as cysD, bcaP, dppA, hisC, gltA, rpsE, purL, amtB as well as a number of hypothetical genes, pseudogenes and notably genetic elements located on the non-coding strand of annotated open reading frames. Furthermore genes that are likely to be involved in interactions with bacteria used in the mixed strain starter culture were identified.

  18. [Production and partial characterization of beta-galactosidase from Kluyveromyces lactis grown in deproteinized whey].

    Science.gov (United States)

    Ramírez Matheus, Alejandra O; Rivas, Nilo

    2003-06-01

    The purpose of this work was to optimize the beta-galactosidase production by Kluyveromyces lactis, applying the Surface Response Methodology (SRM) and using deproteinized whey as fermentation medium. An Orthogonal Central Compound Design (OCCD) was used without repetition, with four factors: temperature, pH, agitation speed and fermentation time. Then, enzyme activity (U/ml) as response variable was used. Thirty trials in twenty-five treatments, with six repetitions at the central point, were carried out, in a New Brunswick Bioflo 2000 fermentor with a volume of 2 liters. The deproteinized whey obtained by thermocoagulation was chemically analyzed. The results were: moisture 93.83%, total solids 6.17%, protein 0.44%, lactose 4.85%, acidity 0.43% and pH 4.58. The best conditions in the enzyme production were: temperature 30.3 degrees C, pH 4.68, agitation speed 191 r.p.m. and fermentation time 18.5 h. with an enzyme production of 8.3 U/ml. The degree of purification obtained was 7.4 times and the yield was 50.8%. The purified enzyme had an optimum temperature of 60 degrees C and a pH of 6.2. This work shows that the yeast Kluyveromyces lactis grown in deproteinized whey is able to produce the enzyme beta-galactosidase and SRM can be used in the fermentology processes, specifically in determining the best suitable operation conditions.

  19. Statistical investigation of Kluyveromyces lactis cells permeabilization with ethanol by response surface methodology

    Directory of Open Access Journals (Sweden)

    Janaína T. de Faria

    2013-12-01

    Full Text Available The aim of our study was to select the optimal operating conditions to permeabilize Kluyveromyces lactis cells using ethanol as a solvent as an alternative to cell disruption and extraction. Cell permeabilization was carried out by a non-mechanical method consisting of chemical treatment with ethanol, and the results were expressed as β-galactosidase activity. Experiments were conducted under different conditions of ethanol concentration, treatment time and temperature according to a central composite rotatable design (CCRD, and the collected results were then worked out by response surface methodology (RSM. Cell permeabilization was improved by an increase in ethanol concentration and simultaneous decreases in the incubation temperature and treatment time. Such an approach allowed us to identify an optimal range of the independent variables within which the β-galactosidase activity was optimized. A maximum permeabilization of 2,816 mmol L-1 oNP min-1 g-1 was obtained by treating cells with 75.0% v/v of ethanol at 20.0 °C for 15.0 min. The proposed methodology resulted to be effective and suited for K. lactis cells permeabilization at a lab-scale and promises to be of possible interest for future applications mainly in the food industry.

  20. Identification and functional characterisation of cellobiose and lactose transport systems in Lactococcus lactis IL1403.

    Science.gov (United States)

    Kowalczyk, Magdalena; Cocaign-Bousquet, Muriel; Loubiere, Pascal; Bardowski, Jacek

    2008-03-01

    Physiological, biochemical and macroarray analyses of Lactococcus lactis IL1403 and its ccpA and bglR single and double mutants engaged in lactose and beta-glucosides catabolism were performed. The kinetic analysis indicated the presence of different transport systems for salicin and cellobiose. The control of salicin catabolism was found to be mediated by the transcriptional regulator BglR and the CcpA protein. The transcriptional analysis by macroarray technology of genes from the PEP:PTS regions showed that several genes, like ybhE, celB, ptcB and ptcA, were expressed at higher levels both in wild type cells exposed to cellobiose and in the ccpA mutant. We also demonstrated that in L. lactis IL1403 cultured on medium with cellobiose and lactose as carbon sources, after the first phase of cellobiose consumption and then co-metabolism of the two sugars, when cellobiose is exhausted the strain uses lactose as the only carbon source. These data could indicate that lactose and cellobiose are transported by a unique system-a PTS carrier induced by the presence of cellobiose, and negatively controlled by the CcpA regulator.

  1. Statistical investigation of Kluyveromyces lactis cells permeabilization with ethanol by response surface methodology

    Science.gov (United States)

    de Faria, Janaína T.; Rocha, Pollyana F.; Converti, Attilio; Passos, Flávia M.L.; Minim, Luis A.; Sampaio, Fábio C.

    2013-01-01

    The aim of our study was to select the optimal operating conditions to permeabilize Kluyveromyces lactis cells using ethanol as a solvent as an alternative to cell disruption and extraction. Cell permeabilization was carried out by a non-mechanical method consisting of chemical treatment with ethanol, and the results were expressed as β-galactosidase activity. Experiments were conducted under different conditions of ethanol concentration, treatment time and temperature according to a central composite rotatable design (CCRD), and the collected results were then worked out by response surface methodology (RSM). Cell permeabilization was improved by an increase in ethanol concentration and simultaneous decreases in the incubation temperature and treatment time. Such an approach allowed us to identify an optimal range of the independent variables within which the β-galactosidase activity was optimized. A maximum permeabilization of 2,816 mmol L−1 oNP min−1 g−1 was obtained by treating cells with 75.0% v/v of ethanol at 20.0 °C for 15.0 min. The proposed methodology resulted to be effective and suited for K. lactis cells permeabilization at a lab-scale and promises to be of possible interest for future applications mainly in the food industry. PMID:24688494

  2. Identification of Bifidobacterium Strains Isolated from Kashk-e Zard: A Traditional Iranian Fermented Cereal-Dairy Based Food

    Directory of Open Access Journals (Sweden)

    Mashak

    2016-09-01

    Full Text Available Objectives The genus Bifidobactrium enjoys considerable significance among the probiotic bacteria for having appropriately adapted to the human gastrointestinal tract. As the properties of Bifidobacteria are strain-oriented and niche-dependent, there is growing interest in studying the different sources of these probiotics. Kashk-e Zard, a traditional fermented food produced from wheat and yogurt through a two-week, two-step fermentation process, is rich in probiotics and is worthy of study in this regard. The present study aimed to identify Bifidobacterium spp. in Kashk-e Zard. Methods Twenty-three samples of Kashk-e Zard were collected and subjected to Bifidobacterium identification experiments. Polymerase chain reaction (PCR and sequencing methods were applied for bacterial identification. Results Twelve of the isolates obtained were G +, rod-shaped, and catalase-, whereas only three of them identified positive for fructose 6-phosphate phosphoketolase (F6PPK a Bifidobacterium specific test and mupirocin resistance. These three isolates were then considered for further identification using the 16SrDNA sequencing technique. Conclusions Although carbohydrate fermentation patterns specified these three isolates as B. infantis, B. bifidum, and B. longum, the molecular results did not confirm B. longum, which is still also controversial in the literature. Overall, our results demonstrated that Kashk-e Zard is a rich potential source of probiotic bacteria and further investigations should be undertaken.

  3. The effect of quercetin on genetic expression of the commensal gut microbes Bifidobacterium catenulatum, Enterococcus caccae and Ruminococcus gauvreauii.

    Science.gov (United States)

    Firrman, Jenni; Liu, LinShu; Zhang, Liqing; Arango Argoty, Gustavo; Wang, Minqian; Tomasula, Peggy; Kobori, Masuko; Pontious, Sherri; Xiao, Weidong

    2016-12-01

    Quercetin is one of the most abundant polyphenols found in fruits and vegetables. The ability of the gut microbiota to metabolize quercetin has been previously documented; however, the effect that quercetin may have on commensal gut microbes remains unclear. In the present study, the effects of quercetin on the commensal gut microbes Ruminococcus gauvreauii, Bifidobacterium catenulatum and Enterococcus caccae were determined through evaluation of growth patterns and cell morphology, and analysis of genetic expression profiles between quercetin treated and non-treated groups using Single Molecule RNA sequencing via Helicos technology. Results of this study revealed that phenotypically, quercetin did not prevent growth of Ruminococcus gauvreauii, mildly suppressed growth of Bifidobacterium catenulatum, and moderately inhibited growth of Enterococcus caccae. Genetic analysis revealed that in response to quercetin, Ruminococcus gauvreauii down regulated genes responsible for protein folding, purine synthesis and metabolism. Bifidobacterium catenulatum increased expression of the ABC transport pathway and decreased metabolic pathways and cell wall synthesis. Enterococcus caccae upregulated genes responsible for energy production and metabolism, and downregulated pathways of stress response, translation and sugar transport. For the first time, the effect of quercetin on the growth and genetic expression of three different commensal gut bacteria was documented. The data provides insight into the interactions between genetic regulation and growth. This is also a unique demonstration of how RNA single molecule sequencing can be used to study the gut microbiota.

  4. Role of Bifidobacterium bifidum and plant food extracts in improving microflora and biochemical and cytogenetic parameters in adjuvant arthritis

    Energy Technology Data Exchange (ETDEWEB)

    Al-Okbi, S. Y.; Mohamed, D. A.; Donya, S. M.; Abd El Khalek, A. B.

    2011-07-01

    The aim of the present research was to discover plant food extracts and probiotics that may have bioactivity towards chronic inflammation. Three plant food extract mixtures expected to be rich in phenolic compounds, carotenoids and tocopherols were prepared. The anti-inflammatory activity of the different mixtures as well as probiotic bacteria (Bifidobacterium bifidum) were evaluated in adjuvant arthritis in rats. The anti-inflammatory effect, mechanism of action and safety of the three mixtures and Bifidobacterium bifidum were studied by measuring the size of inflammation and the determination of inflammatory and oxidative stress biomarkers, colonic bacteria profile and specific cytogenetic parameters. The contents of tocopherols, {beta}-carotene and phenolic compounds in the mixtures were determined. The results show that the tested mixtures and Bifidobacterium bifidum possess promising anti-inflammatory effects. The mechanism of action seems to involve a reduction in oxidative stress and inflammatory bio markers and an effect on colonic microflora. Genotoxicity and DNA fragmentation induced by adjuvant arthritis were prevented after supplementation with the tested mixtures. (Author) 61 refs.

  5. Effect of triple viable bifidobacterium combined with mosapride on hemorheology and serum gastrointestinal hormone levels in patients with functional dyspepsia

    Institute of Scientific and Technical Information of China (English)

    Shun-Bin Ding; Ming-Chun Cai; Wen-Li Zhang; Tian-Hua Jiang

    2016-01-01

    Objective:To observe the effect of triple viable bifidobacterium combined with mosapride on hemorheology and serum gastrointestinal hormone levels in patients with functional dyspepsia (FD).Methods: A total of 127 patients with FD were randomly divided into the observation group (67cases) and the control group (62 cases). The control group was given mosapride, the observation group was given triple viable bifidobacterium on the base of the control group. For 2 months, to observe the efficacy and changes of hemorheology [the whole blood viscosity (high and low shear), plasma viscosity] and serum gastrointestinal hormone levels (MTL, NPY, VIP). Results: After treatment, the observation group of the whole blood viscosity (high and low shear), plasma viscosity were decreased significantly (P0.05). There was significantly difference between the two groups (P<0.05); After treatment, serum MTL, NPY were increased and VIP was increased in both groups (P<0.05), and all indexes of the observation group were improved more significant than those of the control group (P<0.05).Conclusions:Efficacy of triple viable bifidobacterium combined with mosapride is more better than single mosapride in the treatment of FD. It may be related to its effects on hemorheology and gastrointestinal hormone.

  6. Classification of Lactococcus lactis cell envelope proteinase based on gene sequencing, peptides formed after hydrolysis of milk, and computer modeling

    DEFF Research Database (Denmark)

    Børsting, Mette Winther; Qvist, K.B.; Brockmann, E.

    2015-01-01

    Lactococcus lactis strains depend on a proteolytic system for growth in milk to release essential AA from casein. The cleavage specificities of the cell envelope proteinase (CEP) can vary between strains and environments and whether the enzyme is released or bound to the cell wall. Thirty-eight L...

  7. Interaction between ArgR and AhrC controls regulation of arginine metabolism in Lactococcus lactis

    NARCIS (Netherlands)

    Larsen, R; Kok, J; Kuipers, OP

    2005-01-01

    The expression of arginine metabolism in Lactococcus lactis is controlled by the two homologous transcriptional regulators ArgR and AhrC. Genome sequence analyses have shown that the occurrence of multiple homologues of the ArgR family of transcriptional regulators is a common feature of many low-G

  8. Two nucleoside uptake systems in Lactococcus lactis: Competition between purine nucleosides and cytidine allows for modulation of intracellular nucleotide pools

    DEFF Research Database (Denmark)

    Martinussen, Jan; Wadskov-Hansen, Steen Lyders Lerche; Hammer, Karin

    2003-01-01

    in Lactococcus lactis were investigated by measuring the uptake of radioactively labeled nucleosides. The K. for for inosine, cytidine, and uridine was determined to be in the micromolar range. Furthermore, it was found that cytidine and inosine are competitive inhibitors of each other, whereas no competition...

  9. Physiological and Regulatory Effects of Controlled Overproduction of Five Cold Shock Proteins of Lactococcus lactis MG1363

    NARCIS (Netherlands)

    Wouters, Jeroen A.; Mailhes, Marielle; Rombouts, Frank M.; Vos, Willem M. de; Kuipers, Oscar P.; Abee, Tjakko

    2000-01-01

    The physiological and regulatory effects of overproduction of five cold shock proteins (CSPs) of Lactococcus lactis were studied. CspB, CspD, and CspE could be overproduced at high levels (up to 19% of the total protein), whereas for CspA and CspC limited overproduction (0.3 to 0.5% of the total pro

  10. STRESS-RESPONSE IN LACTOCOCCUS-LACTIS - CLONING, EXPRESSION ANALYSIS, AND MUTATION OF THE LACTOCOCCAL SUPEROXIDE-DISMUTASE GENE

    NARCIS (Netherlands)

    SANDERS, JW; LEENHOUTS, KJ; HAANDRIKMAN, AJ; VENEMA, G; KOK, J

    1995-01-01

    In an analysis of the stress response of Lactococcus lactis, three proteins that were induced under low pH culture conditions were detected. One of these was identified as the lactococcal superoxide dismutase (SodA) by N-terminal amino acid sequence analysis. The gene encoding this protein, designat

  11. Biochemical Characterization of ThiT from Lactococcus lactis : A Thiamin Transporter with Picomolar Substrate Binding Affinity

    NARCIS (Netherlands)

    Erkens, Guus B.; Slotboom, Dirk Jan

    2010-01-01

    The putative thiamin transporter ThiT from Lactococeus lactis was overproduced in the membrane of lactococcal cells. In vivo transport assays using radiolabeled thiamin demonstrated that ThiT indeed was involved in thiamin transport. The protein was solubilized from the membranes and purified in det

  12. Expression of the pyrG gene determines the pool sizes of CTP and dCTP in Lactococcus lactis

    DEFF Research Database (Denmark)

    Jørgensen, C.M.; Hammer, Karin; Jensen, Peter Ruhdal;

    2004-01-01

    The pyrG gene from Lactococcus lactis encodes CTP synthase (EC 6.4.3.2), an enzyme converting UTP to CTP. A series of strains were constructed with different levels of pyrG expression by insertion of synthetic constitutive promoters with different strengths in front of pyrG. These strains expressed...

  13. AbiV, a Novel Antiphage Abortive Infection Mechanism on the Chromosome of Lactococcus lactis subsp. cremoris MG1363

    DEFF Research Database (Denmark)

    Haaber, Jakob Brandt Borup; Moineau, Sylvain; Fortier, Louis-Charles;

    2008-01-01

    MG1363. This gene was also found to confer phage resistance to L. lactis MG1363 when it was cloned into an expression vector. A subsequent frameshift mutation in the ORF completely eliminated the phage resistance phenotype, confirming that the ORF was necessary for phage resistance. This ORF provided...

  14. Reconstitution of the Leucine Transport System of Lactococcus lactis into Liposomes Composed of Membrane-Spanning Lipids from Sulfolobus acidocaldarius

    NARCIS (Netherlands)

    in t Veld, Geertruida; Elferink, Maria; Driessen, Arnold J.M.; Konings, Wilhelmus

    1992-01-01

    The effect of bipolar tetraether lipids, extracted from the thermophilic archaebacterium Sulfolobus acidocaldarius, on the branched-chain amino acid transport system of the mesophilic bacterium Lactococcus lactis was investigated. Liposomes were prepared from mixtures of monolayer lipids and the bil

  15. Occurrence of nisin Z production in Lactococcus lactis BFE 1500 isolated from wara, a traditional Nigerian cheese product.

    Science.gov (United States)

    Olasupo, N A; Schillinger, U; Narbad, A; Dodd, H; Holzapfel, W H

    1999-12-15

    Screening for bacteriocin production of 500 strains of lactic acid bacteria (LAB) from various African fermented foods resulted in the detection of a bacteriocin producing Lactococcus lactis (BFE 1500) isolated from a dairy product called wara. The bacteriocin inhibited not only the closely related LAB, but also strains of Listeria monocytogenes, Listeria innocua, Clostridium butyricum, Clostridium perfringens, Bacillis cereus and Staphylococcus aureus. It was heat stable even at autoclaving temperature (121 degrees C for 15 min) and was active over a wide pH range (2-10), but highest activity was observed in the lower pH range. The bacteriocin was inactivated by alpha-chymotrypsin and proteinase K, but not by other proteases. Growth kinetic assay indicated stronger growth inhibition by the bacteriocin produced by Lc. lactis BFE 1500 on L. monocytogenes WS 2250 and B. cereus DSM 2301 than with the nisin A producing strain DSM 20729. Polymerase chain reaction indicated the presence of the nisin operon in strain BFE 1500 and sequencing of its structural gene showed that Lc. lactis BFE 1500 produced the natural nisin variant, nisin Z, as indicated by the substitution of asparagine residue instead of histidine at position 27. The genetic determinants for bacteriocin production in strain BFE 1500 are located on a conjugative transposon. The ability of the bacteriocin produced by Lc. lactis BFE 1500 to inhibit a wide range of food-borne pathogens is of special interest for food safety, especially in the African environment with perennial problems of poor food hygiene.

  16. Versatile vector suite for the extracytoplasmic production and purification of heterologous His-tagged proteins in Lactococcus lactis

    NARCIS (Netherlands)

    Neef, Jolanda; Milder, Fin J.; Koedijk, Danny G. A. M.; Klaassens, Marindy; Heezius, Erik C.; van Strijp, Jos A. G.; Otto, Andreas; Becher, Doerte; van Dijl, Jan Maarten; Buist, Girbe

    2015-01-01

    Recent studies have shown that the Gram-positive bacterium Lactococcus lactis can be exploited for the expression of heterologous proteins; however, a versatile set of vectors suitable for inducible extracellular protein production and subsequent purification of the expressed proteins by immobilized

  17. Functional analysis of glycosyltransferase genes from Lactococcus lactis and other gram-positive cocci: complementation,expression, and diversity

    NARCIS (Netherlands)

    Kranenburg, van R.; Vos, H.R.; Swam, van I.I.; Kleerebezem, M.; Vos, de W.M.

    1999-01-01

    Sixteen exopolysaccharide (EPS)-producing Lactococcus lactis strains were analyzed for the chemical compositions of their EPSS and the locations, sequences, and organization of the eps genes involved in EPS biosynthesis. This allowed the grouping of these strains into three major groups, representat

  18. Transcription profiling of interactions between Lactococcus lactis subsp. cremoris SK11 and Lactobacillus paracasei ATCC 334 during Cheddar cheese simulation.

    Science.gov (United States)

    Desfossés-Foucault, Émilie; LaPointe, Gisèle; Roy, Denis

    2014-05-16

    The starter cultures (Lactococcus sp.) and non-starter lactic acid bacteria (mostly Lactobacillus spp.) are essential to flavor development of Cheddar cheese. The aim of this study was to elucidate the transcriptional interaction between Lactococcus lactis subsp. cremoris SK11 and Lactobacillus paracasei ATCC 334 in mixed cultures during simulated Cheddar cheese manufacture (Pearce activity test) and ripening (slurry). Reverse transcription quantitative PCR (RT-qPCR) was used to quantify the expression of 34 genes common to both bacteria and for eight genes specific to either L. lactis subsp. cremoris SK11 or L. paracasei ATCC 334. The multifactorial analysis (MFA) performed on fold change results for each gene revealed that the genes linked to stress, protein and peptide degradation as well as carbohydrate metabolism of L. paracasei ATCC 334 were especially overexpressed in mixed culture with L. lactis subsp. cremoris SK11 during the ripening simulation. For L. lactis subsp. cremoris SK11, genes coding for amino acid metabolism were more expressed during the cheese manufacture simulation, especially in single culture. These results show how complementary functions of starter and NSLAB contribute to activities useful for flavor development.

  19. Kinetic Properties of a Phosphate-Bond-Driven Glutamate-Glutamine Transport System in Streptococcus lactis and Streptococcus cremoris

    NARCIS (Netherlands)

    POOLMAN, B; SMID, EJ; KONINGS, WN

    1987-01-01

    In Streptococcus lactis ML3 and Streptococcus cremoris Wg2 the uptake of glutamate and glutamine is mediated by the same transport system, which has a 30-fold higher affinity for glutamine than for glutamate at pH 6.0. The apparent affinity constant for transport (KT) of glutamine is 2.5 ± 0.3 μM, i

  20. The anaerobic (Class III) ribonucleotide reductase from Lactococcus lactis : Catalytic properties and allosteric regulation of the pure enzyme system

    NARCIS (Netherlands)

    Torrents, Eduard; Buist, Girbe; Liu, Aimin; Eliasson, Rolf; Kok, Jan; Gibert, Isidre; Gräslund, Astrid; Reichard, Peter

    2000-01-01

    Lactococcus lactis contains an operon with the genes (nrdD and nrdG) for a class III ribonucleotide reductase, Strict anaerobic growth depends on the activity of these genes. Both were sequenced, cloned, and overproduced in Escherichia coli, The corresponding proteins, NrdD and NrdG, were purified c

  1. Lytic Infection of Lactococcus lactis by Bacteriophages Tuc2009 and c2 Triggers Alternative Transcriptional Host Responses

    NARCIS (Netherlands)

    Ainsworth, S.; Zomer, A.L.; Mahony, J.; Sinderen, D. van

    2013-01-01

    Here we present an entire temporal transcriptional profile of Lactococcus lactis subsp. cremoris UC509.9 undergoing lytic infection with two distinct bacteriophages, Tuc2009 and c2. Furthermore, corresponding high-resolution whole-phage genome tiling arrays of both bacteriophages were performed thro

  2. Metabolic and Transcriptional Analysis of Acid Stress in Lactococcus lactis, with a Focus on the Kinetics of Lactic Acid Pools

    NARCIS (Netherlands)

    Carvalho, Ana Lucia; Turner, David L.; Fonseca, Luis L.; Solopova, Ana; Catarino, Teresa; Kuipers, Oscar P.; Voit, Eberhard O.; Neves, Ana Rute; Santos, Helena

    2013-01-01

    The effect of pH on the glucose metabolism of non-growing cells of L. lactis MG1363 was studied by in vivo NMR in the range 4.8 to 6.5. Immediate pH effects on glucose transporters and/or enzyme activities were distinguished from transcriptional/translational effects by using cells grown at the opti

  3. Ability of Lactococcus lactis to export viral capsid antigens: a crucial step for development of live vaccines

    NARCIS (Netherlands)

    Dieye, Y.; Hoekman, A.J.W.; Clier, F.; Juillard, V.; Boot, H.J.; Piard, J.C.

    2003-01-01

    Thefood grade bacterium Lactococcus lactis is a potential vehicle for protein delivery in the gastrointestinal tract. As a model, we constructed lactococcal strains producing antigens of infectious bursal disease virus (IBDV). IBDV infects chickens and causes depletion of B-lymphoid cells in the bur

  4. Interaction of Saccharomyces cerevisiae and Lactococcus lactis in the fermentation and quality of artisanal cachaça

    Directory of Open Access Journals (Sweden)

    Fernanda Paula Carvalho

    2014-11-01

    Full Text Available Lactococcus lactis and Saccharomyces cerevisiae in co-culture were evaluated during sugar cane fermentantion for cachaça production. The inocula containing L. lactis UFLA CA 312 and S. cerevisiae UFLA CA 11 were used in the population of approximately 105 CFU mL-1 and 108 CFU mL-1,  respectively. The sugar cane medium plus 1% of yeast extract (SCM was efficient for growth of L. lactis UFLA CA 312 and S. cerevisiae UFLA CA 11 (letter b -Tukey test. In flasks and vats fermentation the growth of UFLA CA 11 was not negatively influenced by L. lactis UFLA CA 312. However, after 19 h of fermentation, bacterial population showed a slight decrease. Considering parameters higher alcohols and aldehydes, cachaça produced by pure culture of S. cerevisiae was similar to cachaça produced by mixed culture. Cachaça produced by mixed culture showed high values of volatile acidity (letter b -Scott-Knott test being characterized by this parameters in the principal component analysis. High percentage of acceptance (81.10% for the attribute aroma was observed in samples from cachaça produced by mixed culture.

  5. Metabolic Engineering of Mannitol Production in Lactococcus lactis: Influence of Overexpression of Mannitol 1-Phosphate Dehydrogenase in Different Genetic Backgrounds

    NARCIS (Netherlands)

    Wisselink, H.W.; Mars, A.E.; Meer, van der P.; Eggink, G.; Hugenholtz, J.

    2004-01-01

    To obtain a mannitol-producing Lactococcus lactis strain, the mannitol 1-phosphate dehydrogenase gene (mtlD) from Lactobacillus plantarum was overexpressed in a wild-type strain, a lactate dehydrogenase(LDH)-deficient strain, and a strain with reduced phosphofructokinase activity. High-performance l

  6. A POSSIBLE CONTRIBUTION OF MESSENGER-RNA SECONDARY STRUCTURE TO TRANSLATION INITIATION EFFICIENCY IN LACTOCOCCUS-LACTIS

    NARCIS (Netherlands)

    VANDEGUCHTE, M; VANDERLENDE, T; KOK, J; VENEMA, G

    1991-01-01

    Gene expression signals derived from Lactococcus lactis were linked to lacZ-fused genes with different 5'-nucleotide sequences. Computer predictions of mRNA secondary structure were combined with lacZ expression studies to direct base-substitutions that could possibly influence gene expression. Muta

  7. Nisin production of Lactococcus lactis N8 with hemin-stimulated cell respiration in fed-batch fermentation system.

    Science.gov (United States)

    Kördikanlıoğlu, Burcu; Şimşek, Ömer; Saris, Per E J

    2015-01-01

    In this study, nisin production of Lactococcus lactis N8 was optimized by independent variables of glucose, hemin and oxygen concentrations in fed-batch fermentation in which respiration of cells was stimulated with hemin. Response surface model was able to explain the changes of the nisin production of L. lactis N8 in fed-batch fermentation system with high fidelity (R(2) 98%) and insignificant lack of fit. Accordingly, the equation developed indicated the optimum parameters for glucose, hemin, and dissolved oxygen were 8 g L(-1) h(-1) , 3 μg mL(-1) and 40%, respectively. While 1711 IU mL(-1) nisin was produced by L. lactis N8 in control fed-batch fermentation, 5410 IU mL(-1) nisin production was achieved within the relevant optimum parameters where the respiration of cell was stimulated with hemin. Accordingly, nisin production was enhanced 3.1 fold in fed-batch fermentation using hemin. In conclusion the nisin production of L. lactis N8 was enhanced extensively as a result of increasing the biomass by stimulating the cell respiration with adding the hemin in the fed-batch fermentation.

  8. Twofold reduction of phosphofructokinase activity in Lactococcus lactis results in strong decreases in growth rate and in glycolytic flux

    DEFF Research Database (Denmark)

    Andersen, Heidi Winterberg; Solem, Christian; Hammer, Karin;

    2001-01-01

    Two mutant strains of Lactococcus lactis in which the promoter of the las operon, harboring pfk, pyk, and ldh, were replaced by synthetic promoters were constructed. These las mutants had an approximately twofold decrease in the activity of phosphofructokinase, whereas the activities of pyruvate...

  9. The riboflavin transporter RibU in Lactococcus lactis : Molecular characterization of gene expression and the transport mechanism

    NARCIS (Netherlands)

    Burgess, CM; Slotboom, DJ; Geertsma, ER; Duurkens, Hinderika; Poolman, B; van Sinderen, D

    2006-01-01

    This study describes the characterization of the riboflavin transport protein RibU in the lactic acid bacterium Lactococcus lactis subsp. cremoris NZ9000. RibU is predicted to contain five membrane-spanning segments and is a member of a novel transport protein family, not described in the Transport

  10. Complete Sequences of Four Plasmids of Lactococcus lactis subsp. cremoris SK11 Reveal Extensive Adaptation to the Dairy Environment†

    Science.gov (United States)

    Siezen, Roland J.; Renckens, Bernadet; van Swam, Iris; Peters, Sander; van Kranenburg, Richard; Kleerebezem, Michiel; de Vos, Willem M.

    2005-01-01

    Lactococcus lactis strains are known to carry plasmids encoding industrially important traits. L. lactis subsp. cremoris SK11 is widely used by the dairy industry in cheese making. Its complete plasmid complement was sequenced and found to contain the plasmids pSK11A (10,372 bp), pSK11B (13,332 bp), pSK11L (47,165 bp), and pSK11P (75,814 bp). Six highly homologous repB-containing replicons were found, all belonging to the family of lactococcal theta-type replicons. Twenty-three complete insertion sequence elements segment the plasmids into numerous modules, many of which can be identified as functional units or containing functionally related genes. Plasmid-encoded functions previously known to reside on L. lactis SK11 plasmids were now mapped in detail, e.g., lactose utilization (lacR-lacABCDFEGX), the proteolytic system (prtM-prtP, pepO, pepF), and the oligopeptide permease system (oppDFBCA). Newly identified plasmid-encoded functions could facilitate the uptake of various cations, while the pabA and pabB genes could be essential for folate biosynthesis. A competitive advantage could be obtained by using the putative flavin adenine dinucleotide-dependent d-lactate dehydrogenase and oxalate:formate antiporter for enhanced ATP synthesis, while the activity of the predicted α-acetolactate decarboxylase may contribute to the formation of an additional electron sink. Various stress response proteins are plasmid encoded, which could enhance strain robustness. A substantial number of these “adaptation” genes have not been described before on L. lactis plasmids. Moreover, several genes were identified for the first time in L. lactis, possibly reflecting horizontal gene transfer. PMID:16332824

  11. Oral administration of Lactococcus lactis subsp. lactis JCM5805 enhances lung immune response resulting in protection from murine parainfluenza virus infection.

    Directory of Open Access Journals (Sweden)

    Kenta Jounai

    Full Text Available When activated by viral infection, plasmacytoid dendritic cells (pDCs play a primary role in the immune response through secretion of IFN-α. Lactococcus lactis subsp. lactis JCM5805 (JCM5805 is a strain of lactic acid bacteria (LAB that activates murine and human pDCs to express type I and type III interferons (IFNs. JCM5805 has also been shown to activate pDCs via a Toll-like receptor 9 (TLR9 dependent pathway. In this study, we investigated the anti-viral effects of oral administration of JCM5805 using a mouse model of murine parainfluenza virus (mPIV1 infection. JCM5805-fed mice showed a drastic improvement in survival rate, prevention of weight loss, and reduction in lung histopathology scores compared to control mice. We further examined the mechanism of anti-viral effects elicited by JCM5805 administration using naive mice. Microscopic observations showed that JCM5805 was incorporated into CD11c+ immune cells in Peyer's patches (PP and PP pDCs were significantly activated and the expression levels of IFNs were significantly increased. Interestingly, nevertheless resident pDCs at lung were not activated and expressions levels of IFNs at whole lung tissue were not influenced, the expressions of anti-viral factors induced by IFNs, such as Isg15, Oasl2, and Viperin, at lung were up-regulated in JCM5805-fed mice compared to control mice. Therefore expressed IFNs from intestine might be delivered to lung and IFN stimulated genes might be induced. Furthermore, elevated expressions of type I IFNs from lung lymphocytes were observed in response to mPIV1 ex vivo stimulation in JCM5805-fed mice compared to control. This might be due to increased ratio of pDCs located in lung were significantly increased in JCM5805 group. Taken together, a specific LAB strain might be able to affect anti-viral immunological profile in lung via activation of intestinal pDC leading to enhanced anti-viral phenotype in vivo.

  12. Analysis of host-inducing proteome changes in bifidobacterium longum NCC2705 grown in Vivo.

    Science.gov (United States)

    Yuan, Jing; Wang, Bin; Sun, Zhongke; Bo, Xin; Yuan, Xitong; He, Xiang; Zhao, Hongqing; Du, Xinying; Wang, Fang; Jiang, Zheng; Zhang, Ling; Jia, Leili; Wang, Yufei; Wei, Kaihua; Wang, Jie; Zhang, Xuemin; Sun, Yansong; Huang, Liuyu; Zeng, Ming

    2008-01-01

    To investigate the molecular mechanisms underlying the adaptation of Bifidobacterium longum to the intestinal tract, we utilized a new model for rabbit intestinal culture of B. longum and reported the changes in proteomic profiles after incubation in the in vivo environment. By 2D-PAGE coupled with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and/or electrospray ionization tandem mass spectrometry (ESI-MS/MS) analyses, proteomic profiles of B. longum strain NCC2705 grown in the in vivo and in vitro environments were compared. Confirmed by semiquantitative RT-PCR, which exhibited at least a 3-fold change or greater, 19 up-regulated proteins, 14 down-regulated proteins, and 4 proteins with mobility changes were identified during intestinal growth. These identified proteins include key stress proteins, metabolism-related proteins, and proteins related to translation. Our results indicate that some useful proteins are expressed at higher levels in cells during intestinal growth. These proteins reflected the adaptation of B. longum NCC2705 to the intestine, such as EF-Tu which contributes to the retention or attachment as a Bifidobacterium adhesin-like factor, bile salt hydrolase (BSH) which might play an important role in the molecular mechanisms for the initial interaction of probiotic with the intestinal environment, and stress proteins which defend B. longum against the action of bile salts and other harmful ingredients of the gastrointestinal tract (GIT). The most striking fact of our observation was that four proteins GlnA1, PurC, LuxS, and Pgk exhibit clear post-translational modification. Western blot (WB) analysis and Pro-Q Diamond staining revealed that substances of the GIT trigger Pgk and LuxS phosphorylation at Ser/Thr residues for bacteria grown in vivo. These proteins were identified for the first time as bifidobacterial phosphoproteins. Our data suggest that the phosphorylated autoinducer-2 production

  13. The Effects of Bifidobacterium breve on Immune Mediators and Proteome of HT29 Cells Monolayers

    Directory of Open Access Journals (Sweden)

    Borja Sánchez

    2015-01-01

    Full Text Available The use of beneficial microorganisms, the so-called probiotics, to improve human health is gaining popularity. However, not all of the probiotic strains trigger the same responses and they differ in their interaction with the host. In spite of the limited knowledge on mechanisms of action some of the probiotic effects seem to be exerted through maintenance of the gastrointestinal barrier function and modulation of the immune system. In the present work, we have addressed in vitro the response of the intestinal epithelial cell line HT29 to the strain Bifidobacterium breve IPLA20004. In the array of 84 genes involved in inflammation tested, the expression of 12 was modified by the bifidobacteria. The genes of chemokine CXCL6, the chemokine receptor CCR7, and, specially, the complement component C3 were upregulated. Indeed, HT29 cells cocultivated with B. breve produced significantly higher levels of protein C3a. The proteome of HT29 cells showed increased levels of cytokeratin-8 in the presence of B. breve. Altogether, it seems that B. breve IPLA20004 could favor the recruitment of innate immune cells to the mucosa reinforcing, as well as the physical barrier of the intestinal epithelium.

  14. Oral treatment with Bifidobacterium longum 51A reduced inflammation in a murine experimental model of gout.

    Science.gov (United States)

    Vieira, A T; Galvão, I; Amaral, F A; Teixeira, M M; Nicoli, J R; Martins, F S

    2015-01-01

    Gout is an acute inflammatory disease characterised by the presence of uric acid crystals in the joint. This event promotes neutrophil infiltration and activation that leads to tissue damage. We investigated here whether the oral administration of the probiotic strain Bifidobacterium longum 5(1A) (BL) could ameliorate monosodium urate crystal (MSU)-induced inflammation in a murine model of gout. Mice received oral administration of BL or saline daily for 7 days and then were injected with MSU in the knee cavity. Treatment with BL significantly alleviated the inflammatory parameters, as seen by reduced hypernociception, reduced neutrophil accumulation in the joint and myeloperoxidase activity in periarticular tissue. There was inhibition of the production of CXCL1 and interleukin(IL)-1β in joints. Levels of the anti-inflammatory cytokine IL-10 were significantly higher in the knee tissue of mice treated with than control mice injected with MSU. In conclusion, oral BL treatment reduced the inflammatory response in an experimental murine model of gout, suggesting it may be useful as an adjuvant treatment in patients with gout.

  15. Plasmacytoid dendritic cells are crucial in Bifidobacterium adolescentis-mediated inhibition of Yersinia enterocolitica infection.

    Science.gov (United States)

    Wittmann, Alexandra; Autenrieth, Ingo B; Frick, Julia-Stefanie

    2013-01-01

    In industrialized countries bacterial intestinal infections are commonly caused by enteropathogenic Enterobacteriaceae. The interaction of the microbiota with the host immune system determines the adequacy of an appropriate response against pathogens. In this study we addressed whether the probiotic Bifidobacterium adolescentis is protective during intestinal Yersinia enterocolitica infection. Female C57BL/6 mice were fed with B. adolescentis, infected with Yersinia enterocolitica, or B. adolescentis fed and subsequently infected with Yersinia enterocolitica. B. adolescentis fed and Yersinia infected mice were protected from Yersinia infection as indicated by a significantly reduced weight loss and splenic Yersinia load when compared to Yersinia infected mice. Moreover, protection from infection was associated with increased intestinal plasmacytoid dendritic cell and regulatory T-cell frequencies. Plasmacytoid dendritic cell function was investigated using depletion experiments by injecting B. adolescentis fed, Yersinia infected C57BL/6 mice with anti-mouse PDCA-1 antibody, to deplete plasmacytoid dendritic cells, or respective isotype control. The B. adolescentis-mediated protection from Yersinia dissemination to the spleen was abrogated after plasmacytoid dendritic cell depletion indicating a crucial function for pDC in control of intestinal Yersinia infection. We suggest that feeding of B. adolescentis modulates the intestinal immune system in terms of increased plasmacytoid dendritic cell and regulatory T-cell frequencies, which might account for the B. adolescentis-mediated protection from Yersinia enterocolitica infection.

  16. Probiotic Bifidobacterium longum alters gut luminal metabolism through modification of the gut microbial community.

    Science.gov (United States)

    Sugahara, Hirosuke; Odamaki, Toshitaka; Fukuda, Shinji; Kato, Tamotsu; Xiao, Jin-zhong; Abe, Fumiaki; Kikuchi, Jun; Ohno, Hiroshi

    2015-01-01

    Probiotics are well known as health-promoting agents that modulate intestinal microbiota. However, the molecular mechanisms underlying this effect remain unclear. Using gnotobiotic mice harboring 15 strains of predominant human gut-derived microbiota (HGM), we investigated the effects of Bifidobacterium longum BB536 (BB536-HGM) supplementation on the gut luminal metabolism. Nuclear magnetic resonance (NMR)-based metabolomics showed significantly increased fecal levels of pimelate, a precursor of biotin, and butyrate in the BB536-HGM group. In addition, the bioassay revealed significantly elevated fecal levels of biotin in the BB536-HGM group. Metatranscriptomic analysis of fecal microbiota followed by an in vitro bioassay indicated that the elevated biotin level was due to an alteration in metabolism related to biotin synthesis by Bacteroides caccae in this mouse model. Furthermore, the proportion of Eubacterium rectale, a butyrate producer, was significantly higher in the BB536-HGM group than in the group without B. longum BB536 supplementation. Our findings help to elucidate the molecular basis underlying the effect of B. longum BB536 on the gut luminal metabolism through its interactions with the microbial community.

  17. Oral administration of Bifidobacterium longum ameliorates influenza virus infection in mice.

    Science.gov (United States)

    Iwabuchi, Noriyuki; Xiao, Jin-Zhong; Yaeshima, Tomoko; Iwatsuki, Keiji

    2011-01-01

    We investigated whether the oral administration of Bifidobacterium longum BB536 could ameliorate influenza virus (IFV) infection in a mice model. Mice were orally administrated BB536 or saline for 2 weeks and then infected with IFV. Orally administered BB536 significantly alleviated symptoms, reduced the loss of body weight, and inhibited viral proliferation in the lungs relative to the control group findings. Histopathological findings in the lungs were improved in the BB536 group compared to control group findings. There was no significant difference in the levels of interleukin-6 (IL-6), interferon-γ (IFN-γ), IL-10 and IL-12p40 in the lungs between the groups, but the levels of IL-6 and IFN-γ were lower (p=0.076, 0.103, respectively) in the BB536 group compared with those of control group. The levels of IL-6 and IL-10 correlated significantly with the values of weight loss, and the levels of IFN-γ correlated with the virus titers in the lungs. These results suggested the potential of the oral administration of BB536 in ameliorating IFV infection and the possible involvement of anti-inflammatory effects of BB536 in the anti-infection effects against IFV.

  18. Bioaccessible Antioxidants in Milk Fermented by Bifidobacterium longum subsp. longum Strains

    Directory of Open Access Journals (Sweden)

    Mérilie Gagnon

    2015-01-01

    Full Text Available Bifidobacterium longum subsp. longum is among the dominant species of the human gastrointestinal microbiota and could thus have potential as probiotics. New targets such as antioxidant properties have interest for beneficial effects on health. The objective of this study was to evaluate the bioaccessibility of antioxidants in milk fermented by selected B. longum subsp. longum strains during in vitro dynamic digestion. The antioxidant capacity of cell extracts from 38 strains, of which 32 belong to B. longum subsp. longum, was evaluated with the ORAC (oxygen radical absorbance capacity method. On the basis of screening and gene sequence typing by multilocus locus sequence analysis (MLSA, five strains were chosen for fermenting reconstituted skim milk. Antioxidant capacity varied among the strains tested (P=0.0009. Two strains of B. longum subsp. longum (CUETM 172 and 171 showed significantly higher ORAC values than the other bifidobacteria strains. However, there does not appear to be a relationship between gene sequence types and antioxidant capacity. The milk fermented by each of the five strains selected (CUETM 268, 172, 245, 247, or PRO 16-10 did not have higher initial ORAC values compared to the nonfermented milk samples. However, higher bioaccessibility of antioxidants in fermented milk (175–358% was observed during digestion.

  19. Viability of Bifidobacterium Pseudocatenulatum G4 after Spray-Drying and Freeze-Drying

    Directory of Open Access Journals (Sweden)

    Stephenie Wong

    2010-04-01

    Full Text Available Viability of Bifidobacterium pseudocatenulatum G4 following spray-drying and freeze-drying in skim milk was evaluated. After spray-drying, the strain experienced over 99% loss in viability regardless of the air outlet temperature (75 and 85 °C and the heat-adaptation temperature (45 and 65 °C, 30 min. The use of heat-adaptation treatment to improve the thermotolerance of this strain was ineffective. On the other hand, the strain showed a superior survival at 71.65%–82.07% after freeze-drying. Viable populations of 9.319–9.487 log10 cfu/g were obtained when different combinations of skim milk and sugar were used as cryoprotectant. However, the addition of sugars did not result in increased survival during the freeze-drying process. Hence, 10% (w/v skim milk alone is recommended as a suitable protectant and drying medium for this strain. The residual moisture content obtained was 4.41% ± 0.44%.

  20. Allergic Patients with Long-Term Asthma Display Low Levels of Bifidobacterium adolescentis.

    Directory of Open Access Journals (Sweden)

    Arancha Hevia

    Full Text Available Accumulated evidence suggests a relationship between specific allergic processes, such as atopic eczema in children, and an aberrant fecal microbiota. However, little is known about the complete microbiota profile of adult individuals suffering from asthma. We determined the fecal microbiota in 21 adult patients suffering allergic asthma (age 39.43 ± 10.98 years old and compare it with the fecal microbiota of 22 healthy controls (age 39.29 ± 9.21 years old using culture independent techniques. An Ion-Torrent 16S rRNA gene-based amplification and sequencing protocol was used to determine the fecal microbiota profile of the individuals. Sequence microbiota analysis showed that the microbial alpha-diversity was not significantly different between healthy and allergic individuals and no clear clustering of the samples was obtained using an unsupervised principal component analysis. However, the analysis of specific bacterial groups allowed us to detect significantly lower levels of bifidobacteria in patients with long-term asthma. Also, in allergic individuals the Bifidobacterium adolescentis species prevailed within the bifidobacterial population. The reduction in the levels on bifidobacteria in patients with long-term asthma suggests a new target in allergy research and opens possibilities for the therapeutic modulation of the gut microbiota in this group of patients.

  1. Effects of Bifidobacterium Breve Feeding Strategy and Delivery Modes on Experimental Allergic Rhinitis Mice.

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    Jian-jun Ren

    Full Text Available Different delivery modes may affect the susceptibility to allergic diseases. It is still unknown whether early intervention with probiotics would counteract this effect.The effect of different delivery modes on immune status and nasal symptoms was investigated on established allergic rhinitis (AR mouse model. In addition, the immunoregulatory effects and mechanisms of different feeding manners with Bifidobacterium breve(B. breve were examined.Live lyophilized B. breve was orally administered to BALB/c mice born via vaginal delivery(VD or cesarean delivery (CD for 8 consecutive weeks, after which they were sensitized by ovalbumin(OVA to establish experimental AR. Nasal symptoms, serum immunoglobulins, cytokines, splenic percentages of CD4(+CD25(+Foxp3(+ regulatory T(Treg cells and nasal eosinophil infiltration were evaluated.Compared with VD mice, mice delivered via CD demonstrated more serious nasal symptoms, higher concentrations of OVA-specific immunoglobulin (Ig E, more nasal eosinophils and lower percentages of splenic CD4(+CD25(+Foxp3(+Treg cells after establishing experimental AR. These parameters were reversed by administering B. breves hortly after birth. However, the effect of B. breve did not differ between different delivery modes.CD aggravates the nasal symptoms of AR mice compared to VD. This is the first report that oral administration of B. breve shortly after birth can significantly alleviate the symptoms of AR mice born via both deliveries, probably via activation of the regulatory capacity of CD4(+CD25(+Foxp3(+Treg cells.

  2. Mechanisms involved in alleviation of intestinal inflammation by bifidobacterium breve soluble factors.

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    Elise Heuvelin

    Full Text Available OBJECTIVES: Soluble factors released by Bifidobacterium breve C50 (Bb alleviate the secretion of pro-inflammatory cytokines by immune cells, but their effect on intestinal epithelium remains elusive. To decipher the mechanisms accounting for the cross-talk between bacteria/soluble factors and intestinal epithelium, we measured the capacity of the bacteria, its conditioned medium (Bb-CM and other Gram(+ commensal bacteria to dampen inflammatory chemokine secretion. METHODS: TNFalpha-induced chemokine (CXCL8 secretion and alteration of NF-kappaB and AP-1 signalling pathways by Bb were studied by EMSA, confocal microscopy and western blotting. Anti-inflammatory capacity was also tested in vivo in a model of TNBS-induced colitis in mice. RESULTS: Bb and Bb-CM, but not other commensal bacteria, induced a time and dose-dependent inhibition of CXCL8 secretion by epithelial cells driven by both AP-1 and NF-kappaB transcription pathways and implying decreased phosphorylation of p38-MAPK and IkappaB-alpha molecules. In TNBS-induced colitis in mice, Bb-CM decreased the colitis score and inflammatory cytokine expression, an effect reproduced by dendritic cell conditioning with Bb-CM. CONCLUSIONS: Bb and secreted soluble factors contribute positively to intestinal homeostasis by attenuating chemokine production. The results indicate that Bb down regulate inflammation at the epithelial level by inhibiting phosphorylations involved in inflammatory processes and by protective conditioning of dendritic cells.

  3. Fermented inulin hydrolysate by Bifidobacterium breve as cholesterol binder in functional food application

    Science.gov (United States)

    Melanie, Hakiki; Susilowati, Agustine; Maryati, Yati

    2017-01-01

    Inulin hydrolysate is a result of inulin hydrolysis by inulinase enzyme of Scopulariopsis sp.-CBS1 fungi isolated from dahlia tuber skin in the formation of fructooligosaccharides (FOS) as dietary fiber. Inulin hydrolysate fermented by Bifidobacterium breve has a potential as cholesterol binder in digestive system due to dietary fiber content in inulin. This study was conducted to evaluate the best cholesterol binding capacity by the variation of lactic acid bacteria (LAB) culture concentration of 10%, 20% and 30% (v/v), respectively. Fermentation process were conducted with inulin hydrolysate concentration of 25% (w/v), skim milk 7,5% (w/v) and various LAB culture concentration at 40 °C for 0, 12, 24, 36 and 48 hours. The results showed that the variation of LAB culture concentrations affect the cholesterol binding ability in fermented inulin hydrolysate. The fermentation process with 10% LAB culture concentration at 40°C for 48 hours resulted in the highest cholesterol binding capacity (CBC) of 13,69 mg/g at pH 7and 14,44 mg/g at pH 2 with composition of total acids of 0,787%, soluble dietary fiber of 0,396%, insoluble dietary fiber of 5,47%, total solids of 14,476%, total sugars of 472,484 mg/mL, reducing sugar of 92 mg/mL and total plate count (TPC) of 7,278 log CFU/mL, respectively.

  4. Oligosaccharide binding proteins from Bifidobacterium longum subsp. infantis reveal a preference for host glycans.

    Directory of Open Access Journals (Sweden)

    Daniel Garrido

    Full Text Available Bifidobacterium longum subsp. infantis (B. infantis is a common member of the infant intestinal microbiota, and it has been characterized by its foraging capacity for human milk oligosaccharides (HMO. Its genome sequence revealed an overabundance of the Family 1 of solute binding proteins (F1SBPs, part of ABC transporters and associated with the import of oligosaccharides. In this study we have used the Mammalian Glycan Array to determine the specific affinities of these proteins. This was correlated with binding protein expression induced by different prebiotics including HMO. Half of the F1SBPs in B. infantis were determined to bind mammalian oligosaccharides. Their affinities included different blood group structures and mucin oligosaccharides. Related to HMO, other proteins were specific for oligomers of lacto-N-biose (LNB and polylactosamines with different degrees of fucosylation. Growth on HMO induced the expression of specific binding proteins that import HMO isomers, but also bind blood group and mucin oligosaccharides, suggesting coregulated transport mechanisms. The prebiotic inulin induced other family 1 binding proteins with affinity for intestinal glycans. Most of the host glycan F1SBPs in B. infantis do not have homologs in other bifidobacteria. Finally, some of these proteins were found to be adherent to intestinal epithelial cells in vitro. In conclusion, this study represents further evidence for the particular adaptations of B. infantis to the infant gut environment, and helps to understand the molecular mechanisms involved in this process.

  5. Growth and final product formation by Bifidobacterium infantis in aerated fermentations.

    Science.gov (United States)

    González, R; Blancas, A; Santillana, R; Azaola, A; Wacher, C

    2004-10-01

    Fermentation conditions were developed to allow Bifidobacterium infantis to grow in the presence of air. Batch fermentations in TPYG medium, starting from anoxic conditions followed by the application of low airflow rates [0.02-0.1 air volume, per liquid media volume, per minute (vvm)], were analyzed for growth, oxygen uptake, and product formation by the bacterium. Under all aerated fermentations, B. infantis showed high aerotolerance, with a maximum oxygen-specific consumption rate of 0.34 mmol oxygen per gram dry cell weight per hour in the presence of 0.06 vvm. Similar growth yields were obtained under oxic and anoxic conditions (0.11-0.13 and 0.11 g dry cell weight per mmol glucose, respectively). Oxygen also influenced metabolite formation since lactate production and its molar relation to acetate increased and formate decreased with aeration rate. Under anoxic conditions, a maximum concentration of 8.1 mM lactate and an acetate/lactate ratio of 3.5:1 were obtained, while under oxic conditions the lactate concentration increased more than two-fold and the acetate/lactate molar ratio decreased to 1.5:1. The possibility of balancing acetate/lactate molar ratios for organoleptic purposes as well as for obtaining good growth under microaerated conditions was demonstrated.

  6. Biochemical Characterization and Complete Conversion of Coenzyme Specificity of Isocitrate Dehydrogenase from Bifidobacterium longum.

    Science.gov (United States)

    Huang, Shi-Ping; Cheng, Hong-Mei; Wang, Peng; Zhu, Guo-Ping

    2016-02-26

    Bifidobacterium longum is a very important gram-positive non-pathogenic bacterium in the human gastrointestinal tract for keeping the digestive and immune system healthy. Isocitrate dehydrogenase (IDH) from B. longum (BlIDH), a novel member in Type II subfamily, was overexpressed, purified and biochemically characterized in detail. The active form of BlIDH was an 83-kDa homodimer. Kinetic analysis showed BlIDH was a NADP⁺-dependent IDH (NADP-IDH), with a 567- and 193-fold preference for NADP⁺ over NAD⁺ in the presence of Mg(2+) and Mn(2+), respectively. The maximal activity for BlIDH occurred at 60 °C (with Mn(2+)) and 65 °C (with Mg(2+)), and pH 7.5 (with Mn(2+)) and pH 8.0 (with Mg(2+)). Heat-inactivation profiles revealed that BlIDH retained 50% of maximal activity after incubation at 45 °C for 20 min with either Mn(2+) or Mg(2+). Furthermore, the coenzyme specificity of BlIDH can be completely reversed from NADP⁺ to NAD⁺ by a factor of 2387 by replacing six residues. This current work, the first report on the coenzyme specificity conversion of Type II NADP-IDHs, would provide better insight into the evolution of NADP⁺ use by the IDH family.

  7. Biochemical Characterization and Complete Conversion of Coenzyme Specificity of Isocitrate Dehydrogenase from Bifidobacterium longum

    Directory of Open Access Journals (Sweden)

    Shi-Ping Huang

    2016-02-01

    Full Text Available Bifidobacterium longum is a very important gram-positive non-pathogenic bacterium in the human gastrointestinal tract for keeping the digestive and immune system healthy. Isocitrate dehydrogenase (IDH from B. longum (BlIDH, a novel member in Type II subfamily, was overexpressed, purified and biochemically characterized in detail. The active form of BlIDH was an 83-kDa homodimer. Kinetic analysis showed BlIDH was a NADP+-dependent IDH (NADP-IDH, with a 567- and 193-fold preference for NADP+ over NAD+ in the presence of Mg2+ and Mn2+, respectively. The maximal activity for BlIDH occurred at 60 °C (with Mn2+ and 65 °C (with Mg2+, and pH 7.5 (with Mn2+ and pH 8.0 (with Mg2+. Heat-inactivation profiles revealed that BlIDH retained 50% of maximal activity after incubation at 45 °C for 20 min with either Mn2+ or Mg2+. Furthermore, the coenzyme specificity of BlIDH can be completely reversed from NADP+ to NAD+ by a factor of 2387 by replacing six residues. This current work, the first report on the coenzyme specificity conversion of Type II NADP-IDHs, would provide better insight into the evolution of NADP+ use by the IDH family.

  8. Oligosaccharide binding proteins from Bifidobacterium longum subsp. infantis reveal a preference for host glycans.

    Science.gov (United States)

    Garrido, Daniel; Kim, Jae Han; German, J Bruce; Raybould, Helen E; Mills, David A

    2011-03-15

    Bifidobacterium longum subsp. infantis (B. infantis) is a common member of the infant intestinal microbiota, and it has been characterized by its foraging capacity for human milk oligosaccharides (HMO). Its genome sequence revealed an overabundance of the Family 1 of solute binding proteins (F1SBPs), part of ABC transporters and associated with the import of oligosaccharides. In this study we have used the Mammalian Glycan Array to determine the specific affinities of these proteins. This was correlated with binding protein expression induced by different prebiotics including HMO. Half of the F1SBPs in B. infantis were determined to bind mammalian oligosaccharides. Their affinities included different blood group structures and mucin oligosaccharides. Related to HMO, other proteins were specific for oligomers of lacto-N-biose (LNB) and polylactosamines with different degrees of fucosylation. Growth on HMO induced the expression of specific binding proteins that import HMO isomers, but also bind blood group and mucin oligosaccharides, suggesting coregulated transport mechanisms. The prebiotic inulin induced other family 1 binding proteins with affinity for intestinal glycans. Most of the host glycan F1SBPs in B. infantis do not have homologs in other bifidobacteria. Finally, some of these proteins were found to be adherent to intestinal epithelial cells in vitro. In conclusion, this study represents further evidence for the particular adaptations of B. infantis to the infant gut environment, and helps to understand the molecular mechanisms involved in this process.

  9. The regulatory effects of Bifidobacterium infantis on the secretomotor activity of the enteric nervous system

    Directory of Open Access Journals (Sweden)

    Najma Javed

    2013-01-01

    Full Text Available Background: Bifidobacterium infantis (BI and other probiotics are non-pathogenic living organisms that have gained increased attention for their possible therapeutic implications on the health of the digestive tract. The mechanisms by which probiotics exert their effects are largely unknown.Aims: This study explored the protective and regulatory effect of oral BI on the enteric nervous system in the 2, 4, 6-trinitrobenzene sulfonic acid-induced colitis rats. Materials and Methods: Electrical field stimulation and chemical stimulation by 5 hydroxytryptamine or serotonin were used to elicit changes in short-circuit current response of the colonic rat tissue. Results: BI-fed colitis rats expressed trends of higher secretomotor activity and revealed signs of decreased macroscopic inflammatory damage when compared to sham-fed colitis rats, suggesting a protective and preventative role of oral BI. Conclusion: These findings may provide additional insights for understanding the prophylactic and therapeutic value of specific probiotics in intestinal inflammatory disorders, offering the possibility of a non-invasive alternative to toxic and immune-compromising drugs.

  10. Antifungal Activity and Aflatoxin Degradation of Bifidobacterium Bifidum and Lactobacillus Fermentum Against Toxigenic Aspergillus Parasiticus

    Science.gov (United States)

    Ghazvini, Roshanak Daie; Kouhsari, Ebrahim; Zibafar, Ensieh; Hashemi, Seyed Jamal; Amini, Abolfazl; Niknejad, Farhad

    2016-01-01

    Food and feedstuff contamination with aflatoxins (AFTs) is a serious health problem for humans and animals, especially in developing countries. The present study evaluated antifungal activities of two lactic acid bacteria (LAB) against growth and aflatoxin production of toxigenic Aspergillus parasiticus. The mycelial growth inhibition rate of A. parasiticus PTCC 5286 was investigated in the presence of Bifidobacterium bifidum PTCC 1644 and Lactobacillus fermentum PTCC 1744 by the pour plate method. After seven days incubation in yeast extract sucrose broth at 30°C, the mycelial mass was weighed after drying. The inhibitory activity of LAB metabolites against aflatoxin production by A. parasiticus was evaluated using HPLC method. B. bifidum and L. fermentum significantly reduced aflatoxin production and growth rate of A. parasiticus in comparison with the controls (p≤0.05). LAB reduced total aflatoxins and B1, B2, G1 and G2 fractions by more than 99%. Moreover, LAB metabolites reduced the level of standard AFB1, B2, G1 and G2 from 88.8% to 99.8% (p≤0.05). Based on these findings, B. bifidum and L. fermentum are recommended as suitable biocontrol agents against the growth and aflatoxin production by aflatoxigenic Aspergillus species. PMID:28077976

  11. Preclinical Immunomodulation by the Probiotic Bifidobacterium breve M-16V in Early Life

    Science.gov (United States)

    Rigo-Adrover, Maria del Mar; Franch, Àngels; Castell, Margarida; Pérez-Cano, Francisco José

    2016-01-01

    This study aimed to investigate the effect of supplementation with the probiotic Bifidobacterium breve M-16V on the maturation of the intestinal and circulating immune system during suckling. In order to achieve this purpose, neonatal Lewis rats were supplemented with the probiotic strain from the 6th to the 18th day of life. The animals were weighed during the study, and faecal samples were obtained and evaluated daily. On day 19, rats were euthanized and intestinal wash samples, mesenteric lymph node (MLN) cells, splenocytes and intraepithelial lymphocytes (IEL) were obtained. The probiotic supplementation in early life did not modify the growth curve and did not enhance the systemic immune maturation. However, it increased the proportion of cells bearing TLR4 in the MLN and IEL, and enhanced the percentage of the integrin αEβ7+ and CD62L+ cells in the MLN and that of the integrin αEβ7+ cells in the IEL, suggesting an enhancement of the homing process of naïve T lymphocytes to the MLN, and the retention of activated lymphocytes in the intraepithelial compartment. Interestingly, B. breve M-16V enhanced the intestinal IgA synthesis. In conclusion, supplementation with the probiotic strain B. breve M-16V during suckling improves the development of mucosal immunity in early life. PMID:27820846

  12. PCR monitoring of Lactobacillus and Bifidobacterium dynamics in fermentations by piglet intestinal microbiota.

    Science.gov (United States)

    Moura, Patrícia; Simões, Fernanda; Gírio, Francisco; Loureiro-Dias, Maria C; Esteves, M Paula

    2007-04-01

    A new group-specific primer (Lact71R), targeting the 16S-23S rDNA intergenic spacer region of Lactobacillus, was tested in its specificity to amplify rDNA of lactobacilli from piglet intestinal origin by polymerase chain reaction (PCR). Lact71R and Lab0677F, a Lactobacillus group-specific primer targeting the 16S rDNA, generated a common amplicon by PCR with DNA from Lactobacillus and Pediococcus reference strains, but not from Weissella strains. Sequence analysis of clones obtained by PCR amplification with Lact71R and Lab0677F and total DNA isolated from the ileal, caecal and colonic contents of one piglet resulted in Lactobacillus and Lactobacillus-like sequences mainly retrieved from intestinal environments. The primer pair was further validated in a culture independent PCR-analysis to monitor broad fluctuations of lactobacilli populations in fructo-oligosaccharides (FOS) fermentations by piglet intestinal microbiota. Bifidobacterium genus-specific primers were also used for PCR titre determination throughout FOS fermentations, in parallel with lactate and short chain fatty acids (SCFA) quantification. Increases between PCR titres were correlated with lactate detection in early stages of fermentation. Based on the obtained results, a simple monitoring PCR approach is proposed, foreseeing its application to the study of the dynamics of specific bacterial populations in complex environments.

  13. Influence of microencapsulation and spray drying on the viability of Lactobacillus and Bifidobacterium strains.

    Science.gov (United States)

    Goderska, Kamila; Czarnecki, Zbigniew

    2008-01-01

    Improved production methods of starter cultures, which constitute the most important element of probiotic preparations, were investigated. The aim of the presented research was to analyse changes in the viability of Lactobacillus. acidophilus and Bifidobacterium bifidum after stabilization (spray drying, liophilization, fluidization drying) and storage in refrigerated conditions for 4 months. The highest numbers of live cells, up to the fourth month of storage in refrigerated conditions, of the order of 10(7) cfu/g preparation were recorded for the B. bifidum DSM 20239 bacteria in which the N-Tack starch for spray drying was applied. Fluidization drying of encapsulated bacteria allowed obtaining a preparation of the comparable number of live bacterial cells up to the fourth month of storage with those encapsulated bacteria, which were subjected to freeze-drying but the former process was much shorter. The highest survivability of the encapsulated L. acidophilus DSM 20079 and B. bifidum DSM 20239 cells subjected to freeze-drying was obtained using skimmed milk as the cryoprotective substance. Stabilization of bacteria by microencapsulation can give a product easy to store and apply to produce dried food composition.

  14. Safety assessment of Bifidobacterium longum J DM301 based on complete genome sequences

    Institute of Scientific and Technical Information of China (English)

    Yan-Xia Wei; Zhuo-Yang Zhang; Chang Liu; Xiao-Kui Guo; Pradeep K Malakar

    2012-01-01

    AIM: To assess the safety of Bifidobacterium longum (B.longum) JDM301 based on complete genome sequences. METHODS: The complete genome sequences of JDM301 were determined using the GS 20 system. Putative virulence factors, putative antibiotic resistance genes and genes encoding enzymes responsible for harmful metabolites were identified by blast with virulence factors database, antibiotic resistance genes database and genes associated with harmful metabolites in previous reports. Minimum inhibitory concentration of 16 common antimicrobial agents was evaluated by E-test. RESULTS: JDM301 was shown to contain 36 genes associated with antibiotic resistance, 5 enzymes related to harmful metabolites and 162 nonspecific virulence factors mainly associated with transcriptional regulation, adhesion, sugar and amino acid transport. B. longum JDM301 was intrinsically resistant tocipro ciprofloxacin,amikacin, gentamicin and streptomycin and susceptible to vancomycin, amoxicillin, cephalothin, chloramphenicol, erythromycin, ampicillin, cefotaxime, rifampicin, imipenemandtrimethoprim and trimethoprim-sulphamethoxazol. JDM301.JDM301 was moderately resistant to bacitracin, while an earlier study showed that bifidobacteria were susceptible to this antibiotic. A tetracycline resistance gene with the risk of transfer was found in JDM301, which needs to be experimentally validated. CONCLUSION: The safety assessment of JDM301 using information derived from complete bacterial genome will contribute to a wider and deeper insight into the safety of probiotic bacteria.

  15. Coculture fermentations of Bifidobacterium species and Bacteroides thetaiotaomicron reveal a mechanistic insight into the prebiotic effect of inulin-type fructans.

    Science.gov (United States)

    Falony, Gwen; Calmeyn, Thomas; Leroy, Frédéric; De Vuyst, Luc

    2009-04-01

    Four bifidobacteria, each representing a cluster of strains with specific inulin-type-fructan degradation capacities, were grown in coculture fermentations with Bacteroides thetaiotaomicron LMG 11262, a strain able to metabolize both oligofructose and inulin. In a medium for colon bacteria with inulin as the sole added energy source, the ability of the bifidobacteria to compete for this substrate reflected phenotypical variation. Bifidobacterium breve Yakult, a strain that was not able to degrade oligofructose or inulin, was outcompeted by B. thetaiotaomicron LMG 11262. Bifidobacterium adolescentis LMG 10734, a strain that could degrade oligofructose (displaying a preferential breakdown mechanism) but that did not grow on inulin, managed to become competitive when oligofructose and short fractions of inulin started to accumulate in the fermentation medium. Bifidobacterium angulatum LMG 11039(T), a strain that was previously shown to degrade all oligofructose fractions simultaneously and to be able to partially break down inulin, was competitive from the beginning of the fermentation, consuming short fractions of inulin from the moment they appeared. Bifidobacterium longum LMG 11047, representing a cluster of bifidobacteria that shared both high fructose consumption and oligofructose degradation rates and were able to perform partial breakdown of inulin, was the dominating strain in a coculture with B. thetaiotaomicron LMG 11262. These observations indicate that distinct subgroups within the large-intestinal Bifidobacterium population will be stimulated by different groups of prebiotic inulin-type fructans, a variation that could be reflected in differences concerning their health-promoting effects.

  16. The Effect of Hyperosmotic Stress and Nitrogen Starvation on Growth and b-Galactosidase Synthesis in Kluyveromyces lactis and Kluyveromyces marxianus

    OpenAIRE

    2008-01-01

    Kluyveromyces lactis and Kluyveromyces marxianus are industrial yeasts widely used in the production of the b-galactosidase enzyme. Biosynthesis of b-galactosidase is controlled by glucose repression. In this study it was demonstrated that the derepression of b-galactosidase biosynthesis in these yeast strains is inhibited by high osmotic stress. It was found that the b-galactosidase activity of K. lactis and K. marxianus remained approximately at the repressed level when these yeast cells we...

  17. Growth kinetics and physiological behavior of co-cultures of Saccharomyces cerevisiae and Kluyveromyces lactis, fermenting carob sugars extracted with whey.

    Science.gov (United States)

    Rodrigues, B; Lima-Costa, M E; Constantino, A; Raposo, S; Felizardo, C; Gonçalves, D; Fernandes, T; Dionísio, L; Peinado, J M

    2016-10-01

    Alcoholic fermentation of carob waste sugars (sucrose, glucose and fructose) extracted with cheese whey, by co-cultures of Saccharomyces cerevisiae and Kluyveromyces lactis has been analyzed. Growth and fermentation of S. cerevisiae in the carob-whey medium showed an inhibition of about 30% in comparison with water-extracted carob. The inhibition of K. lactis on carob-whey was greater (70%) when compared with the whey medium alone, due to osmolarity problems. Oxygen availability was a very important factor for K. lactis, influencing its fermentation performance. When K. lactis was grown alone on carob-whey medium, lactose was always consumed first, and glucose and fructose were consumed afterwards, only at high aeration conditions. In co-culture with S. cerevisiae, K. lactis was completely inhibited and, at low aeration, died after 3 days; at high aeration this culture could survive but growth and lactose fermentation were only recovered after S. cerevisiae became stationary. To overcome the osmolarity and K. lactis' oxygen problems, the medium had to be diluted and a sequential fermentative process was designed in a STR-3l reactor. K. lactis was inoculated first and, with low aeration (0.13vvm), consumed all the lactose in 48h. Then S. cerevisiae was inoculated, consuming the total of the carob sugars, and producing ethanol in a fed-batch regime. The established co-culture with K. lactis increased S. cerevisiae ethanol tolerance. This fermentation process produced ethanol with good efficiency (80g/l final concentration and a conversion factor of 0.4g ethanol/g sugar), eliminating all the sugars of the mixed waste. These efficient fermentative results pointed to a new joint treatment of agro-industrial wastes which may be implemented successfully, with economic and environmental sustainability for a bioethanol industrial proposal.

  18. Control analysis as a tool to understand the formation of the las operon in Lactococcus lactis

    DEFF Research Database (Denmark)

    Købmann, Brian Jensen; Solem, Christian; Jensen, Peter Ruhdal

    2005-01-01

    In Lactococcus lactis the enzymes phosphofructokinase (PFK), pyruvate kinase (PK) and lactate dehydrogenase (LDH) are uniquely encoded in the las operon and we here apply Metabolic Control Analysis to study the role of this organisation. Earlier work showed that LDH at wildtype level has zero...... control on glycolysis and growth rate but high negative control on formate production. We find that PFK and PK have zero control on glycolysis and growth rate at the wildtype enzyme level but both enzymes exert strong positive control on the glycolytic flux at reduced activities. PK has high positive...... control on formate and acetate production, whereas PFK has no control on these fluxes. Decreased expression of the entire las operon resulted in a strong decrease in growth rate and the glycolytic flux; at 53% expression of the las operon the glycolytic flux was reduced to 44% and the flux control...

  19. Biocidal Inactivation of Lactococcus lactis Bacteriophages: Efficacy and Targets of Commonly Used Sanitizers

    Science.gov (United States)

    Hayes, Stephen; Murphy, James; Mahony, Jennifer; Lugli, Gabriele A.; Ventura, Marco; Noben, Jean-Paul; Franz, Charles M. A. P.; Neve, Horst; Nauta, Arjen; Van Sinderen, Douwe

    2017-01-01

    Lactococcus lactis strains, being intensely used in the dairy industry, are particularly vulnerable to members of the so-called 936 group of phages. Sanitization and disinfection using purpose-made biocidal solutions is a critical step in controlling phage contamination in such dairy processing plants. The susceptibility of 36 936 group phages to biocidal treatments was examined using 14 biocides and commercially available sanitizers. The targets of a number of these biocides were investigated by means of electron microscopic and proteomic analyses. The results from this study highlight significant variations in phage resistance to biocides among 936 phages. Furthermore, rather than possessing resistance to specific biocides or biocide types, biocide-resistant phages tend to possess a broad tolerance to multiple classes of antimicrobial compounds. PMID:28210242

  20. Production of recombinant peanut allergen Ara h 2 using Lactococcus lactis

    DEFF Research Database (Denmark)

    Glenting, J.; Poulsen, Lars K.; Kato, K.;

    2007-01-01

    Background: Natural allergen sources can supply large quantities of authentic allergen mixtures for use as immunotherapeutics. However, such extracts are complex, difficult to define, vary from batch to batch, which may lead to unpredictable efficacy and/ or unacceptable levels of side effects....... The use of recombinant expression systems for allergen production can alleviate some of these issues. Several allergens have been tested in high- level expression systems and in most cases show immunereactivity comparable to their natural counterparts. The gram positive lactic acid bacterium Lactococcus...... of the major allergens in peanut have been described. However, for therapeutic usage more information about the individual allergenic components is needed. In this paper we report recombinant production of the Ara h 2 peanut allergen using L. lactis. Results: A synthetic ara h 2 gene was cloned into an L...

  1. Inducible amplification of gene copy number and heterologous protein production in the yeast Kluyveromyces lactis.

    Science.gov (United States)

    Morlino, G B; Tizzani, L; Fleer, R; Frontali, L; Bianchi, M M

    1999-11-01

    Heterologous protein production can be doubled by increasing the copy number of the corresponding heterologous gene. We constructed a host-vector system in the yeast Kluyveromyces lactis that was able to induce copy number amplification of pKD1 plasmid-based vectors upon expression of an integrated copy of the plasmid recombinase gene. We increased the production and secretion of two heterologous proteins, glucoamylase from the yeast Arxula adeninivorans and mammalian interleukin-1beta, following gene dosage amplification when the heterologous genes were carried by pKD1-based vectors. The choice of the promoters for expression of the integrated recombinase gene and of the episomal heterologous genes are critical for the mitotic stability of the host-vector system.

  2. Comparison of the local immune response against Giardia lamblia cyst wall protein 2 induced by recombinant Lactococcus lactis and Streptococcus gordonii.

    Science.gov (United States)

    Lee, Peter; Abdul-Wahid, Aws; Faubert, Gaétan M

    2009-01-01

    Lactococcus lactis and Streptococcus gordonii are lactic acid bacteria (LAB) currently being advocated for use as live antigen delivery vehicles to mucosal sites. Since both vehicles differ in their capability to persist within the small intestine and in their mode of antigen delivery, we sought to compare them to determine which one was superior. In this study, we compared the efficacy of recombinant L. lactis and S. gordonii to stimulate intestinal immune responses against Giardia lamblia cyst wall protein-2 in BALB/c mice. Oral administration of either vector significantly increased the number of CD4(+) T helper and B-cells in the mesenteric lymph nodes (MLN) and Peyer's patches (PP) of immunized animals. Delivery of recombinant CWP2 (rCWP2) by L. lactis stimulated a balanced IFN-gamma/IL-4 response (MLN and PP cells) and a CWP2-specific intestinal IgA antibody response. Alternatively, delivery of rCWP2 by S. gordonii stimulated a higher frequency of IFN-gamma secreting MLN and PP cells, as well as doubling the amount of CWP2-specific intestinal IgA. In challenge studies, L. lactis and S. gordonii reduced cyst output by 71 and 90%, respectively. When compared to each other, S. gordonii-immunized animals shed 65% fewer cysts than their L. lactis-immunized counterparts. Based on these findings, we concluded that S. gordonii was superior to L. lactis as an intestinal vaccine delivery vehicle.

  3. Recombinant Lactococcus lactis Expressing Haemagglutinin from a Polish Avian H5N1 Isolate and Its Immunological Effect in Preliminary Animal Trials.

    Science.gov (United States)

    Szczepankowska, Agnieszka K; Szatraj, Katarzyna; Sałański, Przemysław; Rózga, Agnieszka; Górecki, Roman K; Bardowski, Jacek K

    2017-01-01

    Lactic acid bacteria (LAB) are Gram-positive, nonpathogenic microorganisms that are gaining much interest as antigen producers for development of live vaccine vectors. Heterologous proteins of different origin have been successfully expressed in various LAB species, including Lactococcus lactis. Recombinant L. lactis strains have been shown to induce specific local and systemic immune responses against various antigens. Our study aimed at constructing a L. lactis strain expressing haemagglutinin of a Polish avian H5H1 influenza isolate and examining its effect on animals. Expression of the cloned H5 gene was achieved using the nisin-controlled gene expression system. Detection of the intracellular H5 antigen produced in L. lactis was performed by Western blot analysis and confirmed using mass spectrometry. The potential of L. lactis recombinant cells to induce an immune response was examined by setting up preliminary immunization trials on chickens and mice. Obtained sera were tested for specific antibodies by ELISA assays. The results of these studies are a promising step toward developing a vaccine against the bird flu using Lactococcus lactis cells as bioreactors for efficient antigen production and delivery to the mucosal surface.

  4. Recombinant Lactococcus lactis Expressing Haemagglutinin from a Polish Avian H5N1 Isolate and Its Immunological Effect in Preliminary Animal Trials

    Directory of Open Access Journals (Sweden)

    Agnieszka K. Szczepankowska

    2017-01-01

    Full Text Available Lactic acid bacteria (LAB are Gram-positive, nonpathogenic microorganisms that are gaining much interest as antigen producers for development of live vaccine vectors. Heterologous proteins of different origin have been successfully expressed in various LAB species, including Lactococcus lactis. Recombinant L. lactis strains have been shown to induce specific local and systemic immune responses against various antigens. Our study aimed at constructing a L. lactis strain expressing haemagglutinin of a Polish avian H5H1 influenza isolate and examining its effect on animals. Expression of the cloned H5 gene was achieved using the nisin-controlled gene expression system. Detection of the intracellular H5 antigen produced in L. lactis was performed by Western blot analysis and confirmed using mass spectrometry. The potential of L. lactis recombinant cells to induce an immune response was examined by setting up preliminary immunization trials on chickens and mice. Obtained sera were tested for specific antibodies by ELISA assays. The results of these studies are a promising step toward developing a vaccine against the bird flu using Lactococcus lactis cells as bioreactors for efficient antigen production and delivery to the mucosal surface.

  5. Recombinant Lactococcus lactis Expressing Haemagglutinin from a Polish Avian H5N1 Isolate and Its Immunological Effect in Preliminary Animal Trials

    Science.gov (United States)

    Szatraj, Katarzyna; Sałański, Przemysław; Rózga, Agnieszka; Górecki, Roman K.; Bardowski, Jacek K.

    2017-01-01

    Lactic acid bacteria (LAB) are Gram-positive, nonpathogenic microorganisms that are gaining much interest as antigen producers for development of live vaccine vectors. Heterologous proteins of different origin have been successfully expressed in various LAB species, including Lactococcus lactis. Recombinant L. lactis strains have been shown to induce specific local and systemic immune responses against various antigens. Our study aimed at constructing a L. lactis strain expressing haemagglutinin of a Polish avian H5H1 influenza isolate and examining its effect on animals. Expression of the cloned H5 gene was achieved using the nisin-controlled gene expression system. Detection of the intracellular H5 antigen produced in L. lactis was performed by Western blot analysis and confirmed using mass spectrometry. The potential of L. lactis recombinant cells to induce an immune response was examined by setting up preliminary immunization trials on chickens and mice. Obtained sera were tested for specific antibodies by ELISA assays. The results of these studies are a promising step toward developing a vaccine against the bird flu using Lactococcus lactis cells as bioreactors for efficient antigen production and delivery to the mucosal surface. PMID:28321412

  6. Optimization of the Lactococcus lactis nisin-controlled gene expression system NICE for industrial applications

    Directory of Open Access Journals (Sweden)

    Mond James

    2005-05-01

    Full Text Available Abstract Background The nisin-controlled gene expression system NICE of Lactococcus lactis is one of the most widely used expression systems in Gram-positive bacteria. Despite its widespread use, no optimization of the culture conditions and nisin induction has been carried out to obtain maximum yields. As a model system induced production of lysostaphin, an antibacterial protein (mainly against Staphylococcus aureus produced by S. simulans biovar. Staphylolyticus, was used. Three main areas need optimization for maximum yields: cell density, nisin-controlled induction and protein production, and parameters specific for the target-protein. Results In a series of pH-controlled fermentations the following parameters were optimized: pH of the culture, use of NaOH or NH4OH as neutralizing agent, the addition of zinc and phosphate, the fermentation temperature, the time point of induction (cell density of the culture, the amount of nisin added for induction and the amount of three basic medium components, i.e. yeast extract, peptone and lactose. For each culture growth and lysostaphin production was followed. Lysostaphin production yields depended on all parameters that were varied. In the course of the optimization a three-fold increase in lysostaphin yield was achieved from 100 mg/l to 300 mg/l. Conclusion Protein production with the NICE gene expression system in L. lactis strongly depends on the medium composition, the fermentation parameters and the amount of nisin added for induction. Careful optimization of key parameters lead to a significant increase in the yield of the target protein.

  7. Food Safety: Secretome of Lactococcus lactis and Listeria monocytogenes in competition.

    Directory of Open Access Journals (Sweden)

    Isabella Alloggio

    2015-07-01

    Full Text Available Listeria monocytogenes (LM is a foodborne pathogen responsible of listeriosis. In the spreading of this pathology, milk and dairy products are key reservoir for this pathogen1. Food processing represents one of the major steps that could be linked to LM growth. Inhibition of LM growth through competition of Lactococcus lactis (LAC could represent a solution to this problem. Exoproteome of LM and two different strains of Lactic Acid Bacteria in co-culture have been studied in order to highlight mechanisms of bacterial competition useful to improve food safety. Two different strains of LAC and one strain of LM were cultivated in appropriate medium cultures (BHI, also in competition. Filtrated cultures (SECRETOME were lyophilized and resuspended for proteomics analysis. Shotgun analysis on each secretome was performed on nano UPLC-MS system. Obtained data reveal, during competition, the higher production by LM of moonlighting protein Enolase and Glucose 6 Phosphate isomerase, of Septation ring formation regulator EzrA, involved into cell replication and the lower secretion of Endopeptidase P60. In parallel, L. lactis produced higher amounts of Secreted 45 kDa protein and switched from lantibiotic Nisin A production to Nisin Z production. In competition with LM, LAC strain investigated produce higher amounts of Secreted 45 kDa protein with peptidoglycan lytic activity and the selective secretion of Nisin Z probably to improve lantibiotic solubility in less acidic environment. Next step will be validation of obtained results in dairy products. These results are of interesting to design new strategies of fighting LM as contaminant in food from animal origin.Work supported by Ministry of Health-CCM “Milano EXPO 2015 Project: Garantire la sicurezza alimentare- Valorizzare le produzioni”

  8. Pilus biogenesis in Lactococcus lactis: molecular characterization and role in aggregation and biofilm formation.

    Directory of Open Access Journals (Sweden)

    Virginie Oxaran

    Full Text Available The genome of Lactococcus lactis strain IL1403 harbors a putative pilus biogenesis cluster consisting of a sortase C gene flanked by 3 LPxTG protein encoding genes (yhgD, yhgE, and yhhB, called here pil. However, pili were not detected under standard growth conditions. Over-expression of the pil operon resulted in production and display of pili on the surface of lactococci. Functional analysis of the pilus biogenesis machinery indicated that the pilus shaft is formed by oligomers of the YhgE pilin, that the pilus cap is formed by the YhgD pilin and that YhhB is the basal pilin allowing the tethering of the pilus fibers to the cell wall. Oligomerization of pilin subunits was catalyzed by sortase C while anchoring of pili to the cell wall was mediated by sortase A. Piliated L. lactis cells exhibited an auto-aggregation phenotype in liquid cultures, which was attributed to the polymerization of major pilin, YhgE. The piliated lactococci formed thicker, more aerial biofilms compared to those produced by non-piliated bacteria. This phenotype was attributed to oligomers of YhgE. This study provides the first dissection of the pilus biogenesis machinery in a non-pathogenic Gram-positive bacterium. Analysis of natural lactococci isolates from clinical and vegetal environments showed pili production under standard growth conditions. The identification of functional pili in lactococci suggests that the changes they promote in aggregation and biofilm formation may be important for the natural lifestyle as well as for applications in which these bacteria are used.

  9. Human Gut-Commensalic Lactobacillus ruminis ATCC 25644 Displays Sortase-Assembled Surface Piliation: Phenotypic Characterization of Its Fimbrial Operon through In Silico Predictive Analysis and Recombinant Expression in Lactococcus lactis.

    Directory of Open Access Journals (Sweden)

    Xia Yu

    Full Text Available Sortase-dependent surface pili (or fimbriae in Gram-positive bacteria are well documented as a key virulence factor for certain harmful opportunistic pathogens. However, it is only recently known that these multi-subunit protein appendages are also belonging to the "friendly" commensals and now, with this new perspective, they have come to be categorized as a niche-adaptation factor as well. In this regard, it was shown earlier that sortase-assembled piliation is a native fixture of two human intestinal commensalics (i.e., Lactobacillus rhamnosus and Bifidobacterium bifidum, and correspondingly where the pili involved have a significant role in cellular adhesion and immunomodulation processes. We now reveal that intestinal indigenous (or autochthonous Lactobacillus ruminis is another surface-piliated commensal lactobacillar species. Heeding to in silico expectations, the predicted loci for the LrpCBA-called pili are organized tandemly in the L. ruminis genome as a canonical fimbrial operon, which then encodes for three pilin-proteins and a single C-type sortase enzyme. Through electron microscopic means, we showed that these pilus formations are a surface assemblage of tip, basal, and backbone pilin subunits (respectively named LrpC, LrpB, and LrpA in L. ruminis, and also when expressed recombinantly in Lactococcus lactis. As well, by using the recombinant-piliated lactococci, we could define certain ecologically relevant phenotypic traits, such as the ability to adhere to extracellular matrix proteins and gut epithelial cells, but also to effectuate an induced dampening on Toll-like receptor 2 signaling and interleukin-8 responsiveness in immune-related cells. Within the context of the intestinal microcosm, by wielding such niche-advantageous cell-surface properties the LrpCBA pilus would undoubtedly have a requisite functional role in the colonization dynamics of L. ruminis indigeneity. Our study provides only the second description of a

  10. Research progress on genetically engineered vaccine of Bifidobacterium%双歧杆菌基因工程疫苗研究进展

    Institute of Scientific and Technical Information of China (English)

    高峰; 吴利先

    2009-01-01

    双歧杆菌是人类最早发现的生理性细菌之一,是能在健康人肠道内定植的益生菌.如今,随着分子生物技术的发展,双歧杆菌基因工程疫苗研究日趋受到重视.此文综述了双歧杆菌基因工程疫苗的理论基础、研究进展以及前景.%Bifidobacterium is one of the human first discovered physiological bacteria,and a beneficial bacterium that colonizing in the healthy people's intestinal canal.Today,with the development of molecular biotechnology,people pay more attention to the research on genetically engineered vaccine of Bifidobacterium.In this paper,the basic theory,research progress and the prospect about the genetically engineered vaccine of Bifidobacterium are reviewed.

  11. Modulación de la fisiología gastrointestinal mediante cepas probióticas de Lactobacillus casei y Bifidobacterium bifidum Modulation of gastrointestinal physiology through probiotic strains of Lactobacillus casei and Bifidobacterium bifidum

    OpenAIRE

    J. Barrenetxe; P. Aranguren; A. Grijalba; J.M. Martínez-Peñuela; F. Marzo; Urdaneta, E. (Elena)

    2006-01-01

    En el contexto de la alimentación y la promoción de la salud se sitúan los productos denominados alimentos funcionales que tienen diversos efectos beneficiosos en el organismo, además de los meramente nutricionales. Dentro de estos alimentos funcionales, entre otros, podemos distinguir entre compuestos probióticos y prebióticos. Los microorganismos más utilizados en alimentos probióticos pertenecen a los géneros Lactobacillus y Bifidobacterium. En el presente trabajo se ha estudiado el efecto...

  12. Initial Study on Growth Morphology of Bifidobacterium Filaments%双歧杆菌丝状体生长形态初探

    Institute of Scientific and Technical Information of China (English)

    刘艳霞; 刘蕾; 刘俊康; 邓渝

    2012-01-01

    The changes of the growth and proliferation patterns of bifidobacterium under specific culture conditions were studied. Different concentrations of VB6 and VE nicotinate of drug-sensitive paper were adopted to jointly affect the growth of bifidobacterium. The growth and proliferation of bifidobacterium were observed through scanning atomic force microscope after Cram staining. The results showed that under joint affection of VB, and VE, a phenomenon of bifidobacterium filaments cluster emerged, and the bifid proliferation pattern was inhibited. The cluster growth and proliferation of bifidobacterium could maintain in the first transferred culture, but after three consecutive transferred cultures biiidobacterium resumed to normal proliferation pattern. Therefore, different concentrations of VB6 and VE nicotinate drug sensitive paper could jointly induce bifidobacterium filaments.%研究双歧杆菌在特定培养条件下生长及增殖模式的变化.采用不同浓度维生素B6和维生素E烟酸酯药敏纸片联合影响双歧杆菌生长,通过革兰染色和原子力显微镜扫描观察双歧杆菌的生长、增殖情况.在维生素B6和维生素E联合作用下,双岐杆菌出现丝状体群集生长的现象,且二分裂增殖方式受到抑制.第1次转种于普通培养基中可保持此种生长增殖方式,但连续转种3次后会恢复常规生长增殖方式.不同浓度维生素B6和维生素E烟酸酯药敏纸片联合可诱生双歧杆菌丝状体.

  13. GABA production and structure of gadB/gadC genes in Lactobacillus and Bifidobacterium strains from human microbiota.

    Science.gov (United States)

    Yunes, R A; Poluektova, E U; Dyachkova, M S; Klimina, K M; Kovtun, A S; Averina, O V; Orlova, V S; Danilenko, V N

    2016-12-01

    Gamma-amino butyric acid (GABA) is an active biogenic substance synthesized in plants, fungi, vertebrate animals and bacteria. Lactic acid bacteria are considered the main producers of GABA among bacteria. GABA-producing lactobacilli are isolated from food products such as cheese, yogurt, sourdough, etc. and are the source of bioactive properties assigned to those foods. The ability of human-derived lactobacilli and bifidobacteria to synthesize GABA remains poorly characterized. In this paper, we screened our collection of 135 human-derived Lactobacillus and Bifidobacterium strains for their ability to produce GABA from its precursor monosodium glutamate. Fifty eight strains were able to produce GABA. The most efficient GABA-producers were Bifidobacterium strains (up to 6 g/L). Time profiles of cell growth and GABA production as well as the influence of pyridoxal phosphate on GABA production were studied for L. plantarum 90sk, L. brevis 15f, B. adolescentis 150 and B. angulatum GT102. DNA of these strains was sequenced; the gadB and gadC genes were identified. The presence of these genes was analyzed in 14 metagenomes of healthy individuals. The genes were found in the following genera of bacteria: Bacteroidetes (Bacteroides, Parabacteroides, Alistipes, Odoribacter, Prevotella), Proteobacterium (Esherichia), Firmicutes (Enterococcus), Actinobacteria (Bifidobacterium). These data indicate that gad genes as well as the ability to produce GABA are widely distributed among lactobacilli and bifidobacteria (mainly in L. plantarum, L. brevis, B. adolescentis, B. angulatum, B. dentium) and other gut-derived bacterial species. Perhaps, GABA is involved in the interaction of gut microbiota with the macroorganism and the ability to synthesize GABA may be an important feature in the selection of bacterial strains - psychobiotics.

  14. Physical and physicochemical stability evaluation of cosmetic formulations containing soybean extract fermented by Bifidobacterium animalis

    Directory of Open Access Journals (Sweden)

    Rafael Pinto Vieira

    2009-09-01

    Full Text Available Peel off facial masks, based on polyvinyl alcohol (PVA, are formulations that, after application and drying, form an occlusive film over the face. After removing, they provide cleanness, tensor and moisturizing effects, removing dead cells, residues and other materials deposited on the stratum corneous. The soybean extract fermented by Bifidobacterium animalis has sugars, amino acids, peptides, proteins and free isoflavonoids in high concentrations, when compared to the unfermented extract, providing benefits to the cosmetic formulations like anti-aging effect, moisture, tensor action and emollience. The cosmetic bases of peel off facial masks, added with 5.0% w/w of fermented soybean extract, were submitted to Preliminary and Accelerated Stability Studies. Eight (8 preparations were evaluated in several conditions of temperature (-10.0, 5.0, 22.0 and 45.0 ºC and time (maximum of 15 days, comparing the results with the initial condition (48 h after preparation. The variables observed were: organoleptic characteristics, pH and appearing viscosity value and film drying time. The preparation containing 17.0% w/w of PVA and 0.5% w/w of guar gum was selected between the eight preparations initially prepared, because it presented the best performance in the stability test, being recommended storage at low temperatures (5.0 ºC.As máscaras faciais peel off a base de álcool polivinílico (PVA são formulações que, após a aplicação e secagem, formam um filme oclusivo sobre a face e, após sua remoção, conferem limpeza, ação tensora e hidratação à pele, retirando células mortas do estrato córneo, resíduos e outros materiais depositados. O extrato de soja fermentado por Bifidobacterium animalis possui açúcares, aminoácidos, peptídeos, e alto teor de isoflavonas na forma livre, quando comparado ao leite não fermentado, propiciando benefícios às formulações cosméticas, como ação antienvelhecimento, hidratação, efeito tensor e

  15. Increased amount of Bifidobacterium thermacidophilum and Megasphaera elsdenii in the colonic microbiota of pigs fed a swine dysentery preventive diet containing chicory roots and sweet lupine

    DEFF Research Database (Denmark)

    Mølbak, Lars; Thomsen, L.E.; Jensen, Tim Kåre

    2007-01-01

    . Methods and Results: Terminal restriction fragment length polymorphism (T-RFLP) was used to describe differences in the total colonic microbiota as well as in the populations of Bifidobacterium spp. in pigs fed with a fructan-rich diet and a diet containing resistant carbohydrates. The fructan-rich diet...... has previously been shown to prevent swine dysentery caused by Brachyspira hyodysenteriae. The T-RFLP profiling, 16S rRNA gene cloning and in situ hybridization showed that the pigs fed with the fructan-rich diet had a higher proportion of Bifidobacterium thermacidophilum subsp. porcinum...

  16. 婴儿肠道双歧杆菌种群多样性分析%Analysis of the probiotics Bifidobacterium community in infant intestinal flora

    Institute of Scientific and Technical Information of China (English)

    张和春; 倪新华; 杨国; 王劲松; 范小兵; 杭晓敏

    2006-01-01

    使用分子生物学方法,对4例出生时间为17~120 h婴儿的肠道双歧杆菌种群多样性进行分析.肠道中剖腹产婴儿在出生17 h后,双歧杆菌就已定植;另外,婴儿肠道中双歧杆菌种群有短双歧杆菌(Bifidobacterium breve)和长双歧杆菌(Bifidobacterium longum),分别占双歧杆菌种群的65%和35%.

  17. Oral administration of an immunostimulatory DNA sequence from Bifidobacterium longum improves Th1/Th2 balance in a murine model.

    Science.gov (United States)

    Takahashi, Noritoshi; Kitazawa, Haruki; Iwabuchi, Noriyuki; Xiao, Jin-Zhong; Miyaji, Kazuhiro; Iwatsuki, Keiji; Saito, Tadao

    2006-08-01

    We have reported the antiallergic activities of the immunostimulatory oligodeoxynucleotide (ODN) BL07S, identified from genomic DNA of Bifidobacterium longum BB536 from in vitro and in vivo studies. The present study evaluated the efficiency of ODN BL07S in preventing allergic responses by oral administration. Oral administration of BL07S suppressed serum ovalbumin (OVA)-specific immunoglobulin (Ig) E levels and improved the OVA-specific IgG2a/IgG1 ratio. ODN BL07S increased Th1 cytokine and decreased Th2 cytokine production in splenocytes. These results suggest that immunostimulatory ODNs are potentially associated with the antiallergic effects of probiotics.

  18. Effect of repeated oral administration of Bifidobacterium longum BB536 on apomorphine-induced rearing behavior in mice.

    Science.gov (United States)

    Orikasa, Shuzo; Nabeshima, Kazumi; Iwabuchi, Noriyuki; Xiao, Jin-Zhong

    2016-01-01

    Schizophrenia is a chronic psychiatric illness. Disruption of the dopaminergic system has been suggested to be the pathogenic cause of this disease. The effect of Bifidobacterium longum BB536 (BB536) on schizophrenic behavior was investigated in an animal model. Daily administration of BB536 (10(9) CFU/mouse, p.o. for 2 weeks) was found to reduce rearing behavior augmented by the dopamine receptor agonist apomorphine and to decrease the resting level of plasma corticosterone and the ratio of kynurenine to tryptophan. These results suggest the potential of BB536 for supplemental treatment of the symptoms of schizophrenia.

  19. [Construction and identification of the Bifidobacterium expression system pGEX-TSOL18/B. longum of Taenia solium].

    Science.gov (United States)

    Zhou, Bi-Ying; Liu, Mei-Chen; He, Li-Fang

    2014-06-01

    The TSOL18 gene of Taenia solium was synthesized and cloned into Escherichia coli-Bifidobacteria shuttle vector pGEX-1lambdaT. The recombinant plasmid pGEX-TSOL18 was transformed into Bifidobacterium longum with electroporation. The recombinant plasmid containing TSOL18 gene was identified by restriction endonuclease analysis, PCR and DNA sequencing. The length of synthesized TSOL18 gene was 393 bp. The results indicated that the Bifidobacteria expression system pGEX-TSOL18/B. longum was successfully constructed.

  20. The putrescine biosynthesis pathway in Lactococcus lactis is transcriptionally regulated by carbon catabolic repression, mediated by CcpA.

    Science.gov (United States)

    Linares, Daniel M; del Río, Beatriz; Ladero, Victor; Redruello, Begoña; Martín, María Cruz; Fernández, María; Alvarez, Miguel A

    2013-07-01

    Lactococcus lactis is the lactic acid bacterium most widely used by the dairy industry as a starter for the manufacture of fermented products such as cheese and buttermilk. However, some strains produce putrescine from agmatine via the agmatine deiminase (AGDI) pathway. The proteins involved in this pathway, including those necessary for agmatine uptake and conversion into putrescine, are encoded by the aguB, aguD, aguA and aguC genes, which together form an operon. This paper reports the mechanism of regulation of putrescine biosynthesis in L. lactis. It is shown that the aguBDAC operon, which contains a cre site at the promoter of aguB (the first gene of the operon), is transcriptionally regulated by carbon catabolic repression (CCR) mediated by the catabolite control protein CcpA.